Agenda • 5 Days of Stem Cells - a virtual event
A Virtual Event
5 Days of Stem Cells
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Join us for the world’s leading virtual stem cell event.

Agenda

This year’s event features over 30 presentations from thought leaders and prominent researchers around the world. Join them as they share developments, discoveries, and cutting-edge content connected to a variety of stem cell applications, including disease and cell modeling, cell and gene therapy, and much more.

Don’t miss your opportunity to hear from this phenomenal speaker lineup!

The final agenda will be updated frequently over the coming weeks. Please check back for the latest details.

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Jiayin Yang

Chief Technology Officer, Cell Inspire Pharma

Jiayin Yang is currently the Chief Technology Officer of Cell Inspire Pharma, a start-up company in Shenzhen, China that aims to develop candidate therapies for neurodegenerative diseases and rare diseases using stem cell-based models. He received his Ph.D. in stem cells and regenerative medicine from the University of Hong Kong. Dr. Yang has 12 years of research experiences in human pluripotent stem cells (PSCs) and genome editing with custom designed nucleases. In combination with these two cutting-edge technologies, Dr. Yang has generated a series of in vitro models with genome engineered induced pluripotent stem cells (iPSCs) for disease models and drug screening.


iPSC-Based High Throughput Platforms for Screening Novel Therapeutics for Treatment of Alzheimer's Disease and Parkinson’s Disease

Abstract

Alzheimer’s disease (AD) and Parkinson’s disease (PD) are complex neurodegenerative diseases that affect millions of people worldwide and are currently no cure. Lacking of suitable cellular models and the inadequate diversity of the existing compound libraries are two hurdles for drug screening for both diseases. To overcome those limitations, we have created novel cellular models based on genetically modified and/or patient-derived human induced pluripotent stem cells (iPSCs). For AD cellular modeling, we site-specifically integrated AD-related genes at the AAVS1 site of a normal iPSC line and created two iPSC lines respectively. Neurons derived from both engineered lines displayed AD phenotypes in vitro, such as significantly elevated Aβ level and p-tau level compared to those of their isogenic parental line. For PD modeling, we obtained iPSC from a patient with young-onset PD (YOPD) who carried multiple mutations in both PARK2 and HTRA2 genes. Neurons derived from the YOPD iPSC displayed typical PD phenotype and severe neurodegenerative features. In order to create high throughput platforms suitable for drug screening, we have also established standardized processes for production of large quantities of uniform neurons from AD and PD iPSC lines and developed robust human neuron-based assays for drug candidate screening.

To test the suitability and robustness of our novel drug screening platforms, we have conducted a preliminary screening of more than one hundred natural chemical compounds plus some herb extracts derived from traditional Chinese medicines (TCM) in a 96-well plate format. Out of the samples screened, we identified more than 20 compounds that showed significant protection for neurons displaying AD and PD phenotypes. Among those 20 positive hits, 5 compounds also significantly downregulate Aβ42 level in the AD model. Our results indicate that combinations of suitable cellular models and the diversity of natural compounds derived from TCM offers a golden opportunity for development of novel therapeutics for treatment of AD and PD. This presentation will discuss details on the generation of our AD and PD cellular models, development of neuron-based assays and potential applications of those novel high throughput screening platforms.

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Julien Muffat

Synthetic Neuroimmunology and Stem Cell Bioengineering, University of Toronto

Dr. Muffat holds the Canada Research Chair in Synthetic Neuroimmunology and Stem Cell Bioengineering. He is a Scientist in the Neurosciences and Mental Health Program at The Hospital for Sick Children, an Assistant Professor in the department of Molecular Genetics, and an Investigator of the Medicine by Design Initiative at the University of Toronto. Dr. Muffat is a graduate of the Biochemistry and Bioengineering department of the Ecole Normale Superieure in Paris-Saclay. He completed a fellowship on the biochemistry of Alzheimer disease at Harvard Medical School, and received his Ph.D. in Molecular and Cellular Neurosciences from Caltech, where he studied neurological aging under the guidance of Seymour Benzer. He pursued his post-doctoral training with Rudolf Jaenisch at the Whitehead Institute at MIT, where he established an in vitro method of differentiation and 3D culture of human iPS cells into microglia-like cells, to model inflammatory etiologies in CNS disorders. He came to Toronto, Canada, in 2018, where his laboratory studies the interactions of the human nervous and innate immune systems, using in vitro approaches.


Personalized Immuno-Neuroscience and Glio-Immunology: Towards Human Culture Models of Innate Responses In The Brain

Abstract

The brain harbors a resident innate immune system, finding its origins early in development. These immune cells, the microglia, along with rare patrolling peripheral blood cells, contribute to the development, maturation, homeostasis, and perhaps eventual demise of the nervous system. We use humanized in vitro models to investigate the role of these cells in health and disease. Mutations and polymorphisms affecting genes expressed solely in microglia are associated with conditions ranging from psychiatric disorders of developmental origin (e.g. Schizophrenia), to age-related neurological disorders (e.g. Alzheimer’s). Using primary benchmarks and single cell profiling, combined with tissue engineering, we aim to better replicate cellular identity in our cultures, as a prerequisite for identification of disease-associated phenotypes. Functional variations affecting microglial homeostatic behavior and reactivity, at different stages of development and different ages, eventually lead to neuronal dysfunction, loss of connectivity, or death. We use the CRISPR/Cas9 toolkit to engineer putative disease-associated variants in pluripotent stem cells. Glial cells such as microglia and astrocytes are capable of sensing and reacting to various stimuli, such as immune/inflammatory modulators. As such, they are uniquely suited to integrate peripheral inputs (pathogens, microbiota, cytokines), as well CNS-centric stimuli (synaptic plasticity, electrophysiological activity, metabolic demands, cell death) over time, ultimately affecting the health of our neural networks, for better or worse. While their roles in development are under evolutionary constraint, their actions in disease or during aging can be maladaptive. Our ultimate goal is to identify and control inflammatory processes that precede, sometimes by decades, the largely irreversible neuronal dysfunction seen in patients with neurological and psychiatric disorders.

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Silvia E. Castro Piedra

Biotechnology Research Center, Costa Rica Institute of Technology

Silvia E. Castro Piedra graduated with a Bachelor degree in Biotecnological Engineering and with a Master’s degree in Microbiology from the University of Costa Rica in 2019. She has over ten years of experience in Tissue Engineering and Regenerative Medicine research. Also during the same time has been a lecturer of embryology, Human Physiology, Biology and Anatomy, as well cell culture techniques in different Universities in Costa Rica. She has experience using laboratory animals. She has also has participated in different international events as TERMIS as a speaker and several poster presentations and owns publications. Because of her personal and particular interest she has participated in different congress organization related to health research and worked with different interdisciplinary groups in more than ten projects.


Stem cell therapies for skin regeneration in animal models, Costa Rica experiences

Abstract:

The Tissue Engineering Laboratory (LAINTEC) of the Costa Rican Institute of Technology (TEC), started activities in 2005 aiming to develop cell therapies focused on skin regeneration. Throughout the last fifteen years, this laboratory has grown and the research has diversified, so now it has more than a dozen of multidisciplinary projects focused on regenerative skin therapies, development of three-dimensional models of skin, muscle and bone, evaluation of new biomaterials, among other research lines. One of the most important areas is the development of skin regeneration therapies. As part of the experience, we have worked with two approaches using different types of stem cells: epidermal stem cell (ESC) and adipose derived stem / stromal cell (ASC). First, ESC were isolated, expanded and characterized analyzing cytokeratines 10, 14 and 15 expression. To evaluate the regeneration potential of ESCs, a controlled full thickness skin wound was performed in the interscapular area in an adult rat model and a plasma-based matrix (PBM) with fibroblasts and keratinocytes applied, compared with a PBM with fibroblasts or PBM alone as control. On the other hand ADSC were isolated and the identificated using CD73, CD90, CD105 and CD45 markers. Also, the differentiation potential was analyzed. To evaluate the regeneration using ASC, 5 six-week-old BalbC mice per group were subcutaneously treated with: ASC in saline solution, ASC seeded on a natural scaffold and scaffold with saline solution. A positive and negative control was also performed. After two weeks of treatment, at a macroscopic level, both treatments showed no significant differences in the regeneration rate. However, the ASC treated group, showed more vascularization and a better collagen fibers organization compared with the control groups. Eventhough is necessary more preclinical testing and evidence, these experiments present the first steps taken in Costa Rica towards cellular therapy development as well as strengthening research in regenerative medicine and tissue engineering.

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Jarmon Lees

St Vincent's Institute of Medical Research

Dr. Jarmon Lees completed his PhD at the University of Melbourne in 2017 where he examined the role of metabolism in regulating human pluripotent stem cell pluripotency and neural differentiation. He then worked on the development of a novel pluripotent stem cell growth formulation for the biotech company Vitrolife. In 2018, Jarmon joined the Cardiac Regeneration Group at St Vincent’s as a Research Fellow examining cardiomyopathy in Friedreich’s ataxia and strategies for innervating human cardiac tissue using pluripotent stem cells. The cardiac regeneration lab is involved in modeling a range of heart diseases using a novel vascularised and innervated cardiac organoid model. 


Heart disease in a dish: human iPSC-derived multicellular cardiac organoids  

Abstract:

Cardiovascular disease is the leading cause of death worldwide necessitating accurate human disease models to improve our cellular and molecular understanding of heart diseases and facilitate preclinical trials. Here we have developed an advanced 3D multicellular human heart tissue model for modelling cardiovascular diseases. Human induced pluripotent stem cells (iPSCs) were differentiated into cardiomyocytes, endothelial cells and sympathetic neurons for construction of vascularised and innervated cardiac organoids, which can be maintained for at least 4 weeks in culture. Histological analysis showed CD31+ endothelial networks and tyrosine hydroxylase+ neural networks interspersed throughout the organoids. Single-cell RNAseq showed reproducibility of our cardiac organoids containing all input cell types and cells in an intermediate state. Cardiac organoids exhibited spontaneous and synchronous contractions at ~160 bpm. Subjecting the cardiac organoid to an acute ischaemia-reperfusion injury or chronic hyperglycaemic and hyperlipidaemic (to simulate type 2 diabetes) conditions increased the release of lactate dehydrogenase (an indicator of cell death). Cardiac organoids subjected to chronic hyperglycaemia and hyperlipidaemia also showed a reduction in contraction rate and prolongation in relaxation time. This indicates the capability of the cardiac organoids to simulate cardiac responses to ischaemic heart disease and type 2 diabetes-induced cardiac injury. This in vitro human cardiac tissue will be an ideal pre-clinical human model to study and develop novel therapeutics for heart diseases.

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Laura Steenpass

Technical University Braunschweig

Dr. Laura Steenpass received her PhD at the Children’s Cancer Research Institute (CCRI) in 2002 where her research focus was protein-protein interactions and Ewing’s sarcoma. She was later a PostDoc in the lab of Prof. Denise Barlow at the Center for Molecular Medicine (CeMM) in Vienna, Austria until 2006. Her research focus was genomic imprinting at the murine lgf2r/Airn locus. Dr. Steenpass also served as a PostDoc at the Department of Pediatric Oncology, Hematology and Immunology, University Hospital Düsseldorf, Germany from 2006 to 2009. She now serves as a scientist and PI at the Institute of Human Genetics, University Hospital Essen, Germany with a research focus of modeling of imprinting diseases and retinoblastoma with human pluripotent stem cells. Most recently, Dr. Steenpass is Head of Department Human and Animal Cell Cultures at the Leibniz-Institute DSMZ – German Collection of Microorganisms and Cell Cultures and Professor for Cell Biology at the Technical University Braunschweig, Germany. 


A human stem cell based model for retinoblastoma 

Abstract

Retinoblastoma is the most common eye cancer occurring in children under the age of five. It is caused by biallelic inactivation of the retinoblastoma gene RB1, presumably in cone precursor cells of the human retina. Efforts to model retinoblastoma in mouse are not satisfactory as the mutation of Rb1 alone is not sufficient for tumor formation. In order to analyze especially early stages of retinoblastoma we have created a human organoid-based model using the CRISPR/Cas9 system. The human embryonic stem cell (hESC) line H9 was modified to generate sublines carrying a random mutation in exon 3 (close to the splice donor site) or a deletion of RB1 exon1 including the promoter. Both modifications were established either on one or both RB1 alleles. All cell lines have been characterized thoroughly and tested for pluripotency. Comparative differentiation into retinal organoids using the parental cell line H9, a heterozygous and a homozygous subline of each modification was performed. As analyzed by immunocytochemistry, generated retinal organoids contain all seven retinal cell types – the mature rod (NRL) and cone (ARR3) photoreceptors are present in the outer layer and ganglion cells (POU4F1), Müller glia cells (VIM), amacrine cells (TFAP2A), bipolar cells (PRKCA) and horizontal cells (PROX1) are located in the inner layer. Over time, the neural retina layer of the RB1 homozygous organoids became loose and disorganized compared to RB1 wildtype and RB1 heterozygous organoids. Immunostainings on day 152 indicated enhanced proliferation (Ki67), a decrease in rod photoreceptors and an absence of amacrine cells in RB1-/- organoids. Co-staining for Ki67 and ARR3 and subsequent quantification of cell numbers in microscopy images revealed a significant increase of proliferating ARR3+ cone photoreceptors in RB1-/- organoids. Analysis of mRNA expression of marker genes by qRT-PCR on day 126 supported theRB1-/- specific changes in proliferating and amacrine cells and rod photoreceptors. Based on these data, we are convinced that retinal organoids are a suitable model to enhance studies on development of retinoblastoma.

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Jean Lu

Genomics Research Center, Academia Sinica

Research Interests

We use high throughput screening to target chemicals/cytokines/shRNAs those can efficiently promote cell renewal/differentiation/transdifferentiation

Educational Background

2000 Ph.D. Institute of Microbiology, National Taiwan University

1994 M. S. Institute of Molecular Medicine, National Taiwan University

1992 B.S. Department of Medical Technology, National Taiwan University

Professional Experiences

2018-present Adjunct Associate Professor, Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan.

2016-present Adjunct Associate Professor, Department of Life Science, Tzu Chi University, Taiwan

2015-present Associate Research Fellow, Stem Cell Program, Genomics Research Center, Academia Sinica, Taiwan

2015-present Adjunct Associate Professor, Genomics and System Biology Program, College of Life Science, National Taiwan University, Taiwan

2003-2007 Postdoctoral Fellow/Associate, Molecular, Department of Cellular, and Developmental Biology, Yale University, USA


A high-throughput functional screen reveals human embryonic stem cell self-renewal signals

Abstract

Only very few studies focus on the cytokines secreted by hESCs. We screened and investigated a chemokine (C-X-C motif) ligand 14 (CXCL14), is a downstream effector of ATF1 is critical for ESC renewal. Disruption of CXCL14 expression downregulate the expression of Oct4/Sox2/Nanog, arrest the cell cycle at G0/G1 stage, and further increased the expression levels of differentiated markers. Furthermore, by co-immunoprecipitation, ELISA, and duo-link assay, we demonstrated that CXCL14 is the ligand for the insulin-like growth factor 1 receptor (IGF-1R). CXCL14 can stimulate IGF-1R signal cascade to maintain hESC self-renewal. For now, the literature indicates that all receptors in the CXCL family belong to G protein-coupled receptors (GPCRs). This study is the first to identify that a CXCL chemokine can bind to and trigger a receptor tyrosine kinase (RTK), IGF-1R. In addition to IGF-1 and insulin, this is also the third ligand of IGF1-R. These findings increase our understanding of signal transduction and stem cell biology.

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Thomas Durcan

MNI, McGill University

As an assistant professor at the Montreal Neurological Institute (The Neuro) and McGill University, my research focus is on applying patient-derived stem cells towards the development of phenotypic discovery assays and 3D neuronal organoid models for both neurodegenerative and neurodevelopmental disorders. As associate director of the Early Drug Discovery Unit (EDDU) at The Neuro, I oversee a team of 40+ research staff and students, committed to applying novel stem cell technology, combined with CRISPR genome editing, organoid models and new microfluidic technologies towards elucidating the underlying causes of these complex disorders. Combined with new approaches in the group towards building multiomic profiles and predictive computational models with patient-derived IPSC cells, the long term strategy over the next decade is to identify new personalized precision therapies that can be applied towards building clinical trials on a dish. Further information on the EDDU can be found on our website https://www.mcgill.ca/neuro/open-science/open-science-platforms/eddu 


Modelling disorders of the brain through patient-derived stem cells

Abstract

My research focus is on applying patient-derived stem cells towards the development of phenotypic discovery assays and 3D mini-brain models for both neurodegenerative and neurodevelopmental disorders. As associate director of the Early Drug Discovery Unit (EDDU) at the Montreal Neurological Institute (MNI), I oversee a team of over 35 research staff and students, committed to applying novel stem cell technology, combined with CRISPR genome editing, mini-brain models and new microfluidic technologies towards elucidating the underlying causes of these complex disorders. Combined with new approaches in the group towards building MultiOmics profiles on the patient-derived IPSC cells, the long-term strategy is to identify new personalized precision therapies that can be applied towards building clinical trials on a dish. For my talk, I will discuss our 3D drug discovery pipeline, in which we focus on translating findings from 2D neurons, into 3D brain organoids, and ultimately into lead compounds. I will present work on how we generate these cells, the quality control process and also some of the new technologies we are developing in the group for working with 3D neuronal organoids

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Larisa Haupt

Queensland University of Technology

Associate Professor Larisa Haupt is a Principal Research Fellow and the Neurogenesis and Stem Cell Group Leader and Laboratory Manager within the Genomics Research Centre, and Program Leader in Diagnostics and Functional Genomics within the QUT Tier 1 Centre for Genomics and Personalised Health at IHBI. A/Prof Haupt has extensive research expertise in the extracellular matrix, stem cells, cell and molecular biology and human molecular genetics. Her research team has a particular interest in the role of the extracellular matrix, with a focus on the proteoglycans, in the regulation and dysregulation of cell behaviour including lineage specification, neurodegeneration and cancer. A/Prof Haupt and her team utilise molecular and cell biological two- and three-dimensional human stem cell culture models in conjunction with next generation sequencing platforms to unravel these complex mechanisms in humans. In the last 10 years, A/Prof Haupt has published 50 manuscripts (43 in the last 5 years; 13 to date in 2020), with a current Google Scholar H index of 27 and an i10-index of 62. 


In vitro models of human neurogenesis

Larisa M Haupt, Rachel K. Okolicsanyi, Chieh Yu, Lotta E. Oikari, Ian W Peall, Lyn R. Griffiths

Abstract

Assessing the functionality of neuronal cell types is critical to their efficacy in future applications. Understanding how these processes are regulated will help to further unravel the structural complexity of the human brain, and the role of associated biological and other factors in neurogenesis. This information will also have important ramifications for the development of 2D and 3D models to translate cells toward their successful integration of newly formed neurons into existing/remaining neural circuits. The complexity of the neural niche and the ubiquitous presence of the protetoglycan proteins in the neural microenvironment suggest that this will be achievable not by one assay but will more likely be achieved through a combination of data and approaches to identify specific markers and regulatory pathways. Our approach uses human stem cell models and cell lines including human mesenchymal stem cells (hMSC), embryonic derived human neural stem cells (hNSC H9) and ENStem-A neural progenitor cells, as well as the immortalised human neural progenitors (ReNcell CX cell line), in 2D and developing 3D cultures to examine the role of proteoglycans and their use as markers of lineage potential and specification.

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Igor Slukvin

University of Wisconsin

Dr. Igor Slukvin is a Professor of Pathology and Laboratory Medicine and Cell and Regenerative Biology at the University of Wisconsin, Madison. He received his medical education and PhD degree at Kiev Medical University, Ukraine. After moving to the United States, he completed postdoctoral training and medical residency in pathology at UW Madison and later became the faculty member at the same institution. His research interest is in the understanding of cellular and molecular pathways leading to development of hematopoietic and vascular cells from human pluripotent stem cells (hPSCs). Dr. Slukvin also co-directs Precision Medicine Core at the Wisconsin National Primate Research Center which is focused on establishing next generation animal models and tools for the assessment precision stem cell therapies. His work is relevant for the development of novel sources of cells for bone marrow transplantation, transfusion and cancer and AIDS immunotherapies. He is a cofounder of Cellular Dynamics International and Cynata therapeutics biotechnology companies.


Advancing pluripotent stem cell technologies for research and therapy of blood diseases

Abstract


The derivation of human embryonic stem cells more than 20 years ago by James Thomson at University of Wisconsin followed by advances in cellular reprogramming have created alternative platforms for manufacturing blood cells for transfusion, immunotherapies and transplantation using human pluripotent stem cells (hPSCs). However, development of such therapies depends on our ability to produce the appropriate types of hematopoietic cells in sufficient quantities. Although we have demonstrated the feasibility of generating a variety of blood cell types from hPSCs, significant challenges remain, including de novo generation of hematopoietic stem cells (HSC) and robust production of lymphoid cells from hPSCs. This is due to the limited specification of adult-type definitive hematopoietic progenitors and predominance of myeloid-restricted embryonic hematopoiesis in hPSC differentiation cultures. In the embryo, lymphoid progenitors and hematopoietic stem cells (HSCs) arise from hemogenic endothelium (HE) lining arteries, but not veins. In our recent studies we identified HE in hPSC cultures and demonstrated the important role of NOTCH and arterial signaling in specification of definitive HE, thus providing an innovative strategy to aid in generating of definitive lymphomyeloid progenitors from hPSCs through enhancing arterial programming of HE. In addition, I will discuss our advances in direct blood programming technologies using modified mRNA and the utility of iPSC models for identifying novel factors involved in leukemia stem cell survival.





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Ritu Kumar

Weill Cornell Medicine

Ritu Kumar is an Assistant Professor of Cell and Developmental Biology Research in Surgery at Weill Cornell Medicine, New York, with a strong background in stem cell research. During her Ph.D. She studied the risk factors associated with calcium oxalate urolithiasis. After finishing her Ph.D. in 2005, Ritu joined Mount Sinai School of Medicine (now the Icahn School of Medicine at Mount Sinai), New York as a post-doc. During her post-doc, Ritu investigated molecular mechanisms controlling self-renewal and aging of hematopoietic stem cell compartment. Dr. Kumar joined Weill Cornell Medicine in the year 2009 and her research over the past several years has been focused on understanding the epigenetic basis of somatic cell reprogramming towards induced pluripotent stem cells (iPSC). Ritu discovered, an epigenetic modifier Activation-Induced Cytidine Deaminase (AICDA) is required for the stabilization of mouse iPSCs during reprogramming and even when iPSCs can be stabilized by modifying the levels of transgenes, they fail to achieve naive pluripotent state and are primed for differentiation, hypermethylated and display hyperactive FGF/ERK signaling.  After gaining extensive knowledge on the epigenetic basis of mouse pluripotency, Dr. Kumar has shifted her focus to human pluripotent stem cells and her current research is aimed at to decipher the epigenetic basis of human naïve pluripotency, with the eventual goal to establish a stable population of bona-fide human naive pluripotent stem cells in-vitro, which will eventually benefit various clinical applications of pluripotent stem cells.  Additionally, at the Department of Surgery Dr. Kumar is carrying out multiple projects in, related to patient derived iPSCs for disease modeling and cell replacement therapies purposes, including cardiomyopathies and pancreatic disorders. She is working on a number of collaborative projects with various leading stem cell scientist at WCM and also with researchers at neighboring institutes such as the Memorial Sloan Kettering Cancer Center and Rockefeller University. 

Epigenetic regulation of human naïve pluripotency

Abstract:

Naïve pluripotenct stem cells (PSCs) are the most potent stem cells, with the highest capacity to differentiate into all the cells of the three germ layers without lineage biases, thus hold the greatest promise for the field of regenerative medicine and human disease modeling. Stem cells in pre-implantation epiblast constitutes the naive state, possess unrestricted developmental potency and flexibility to produce all lineages. Conversely, PSCs present in the post implantation blastocyst, designated as epiblast stem cells (EpiSCs) are less plastic and have acquired the genetic and epigenetic landscape for differentiation. That’s why embryonic stem cells (ESCs) are derived from the pre-implantation stage. Contrary to mouse ESCs (mESCs), which are similar to naïve cells, human embryonic stem cells (hESCs) despite their derivation form pre-implantation epiblast are analogous to a late stage EpiSCs and are in primed state. This is a significant drawback for the translational applications of stem cells as primed cells are not at the ground state, are transcriptionally and epigenetically very heterogeneous, contributing to inefficient differentiation and significant variation and non-reproducibility within one and among various cell lines. Epigenetic rewiring is essential for the establishment of naïve pluripotent state. Here I report Activation-Induced Cytidine Deaminase (AICDA) an enzyme involved in DNA-dementylation is a novel regulator of both mouse and human naïve pluripotency. Aicda knockout mPSCs fail to achieve the naïve pluripotent state, and remain primed for differentiation, because of a failure to suppress FGF/ERK signaling. While the mutant cells display marked genomic hypermethylation, suppression of FGF/ERK signaling by AID is independent of deaminase activity. Similarly, AICDA-mutant hESCs also fail to achieve the naïve state and display hyperactive FGF/ERK signaling. Thus, our study identifies AICDA as a novel regulator of naïve pluripotency through its activity on FGF/ERK signaling and DNA-demethylation, which, in future, will help develop strategies to direct human ESCs to a bona-fide naïve state with high efficiency.

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Cláudia Miranda

University of Lisbon

Cláudia Miranda is currently a Post Doctoral Research Fellow at iBB – Institute for Bioengineering and Biosciences/Instituto Superior Técnico.

The main focus of her research is the development of a scalable and cost-effective culture platforms for expansion of human pluripotent stem cells and their controlled differentiation into different lineages for drug discovery and toxicity assessment.

After finishing a Degree in Biochemistry, she received a MSc. in Biotechnology and a Ph.D. in Biotechnology and Biosciences at Instituto Superior Técnico, University of Lisbon, focusing on the 3D culture and neural induction of human pluripotent stem cells.


Development of a robust and scalable culture platform for expansion and neural induction of human pluripotent stem cells in suspension culture

Abstract

The demand for large cell numbers for applications in cellular therapies and drug screening requires the development of scalable platforms capable of generating high quality populations of tissue-specific cells derived from human pluripotent stem cells (hPSCs). This work describes the scaling-up of an aggregate-based suspension culture system for expansion and neural induction of human pluripotent stem cells using a novel medium formulation: Gibco StemScale PSC Suspension Medium.

Here, we studied the ability of StemScale Medium to promote the rapid expansion of hPSC cultures as spheroids grown in suspension. We tested human induced pluripotent stem cell (hiPSC) growth in 6-well plates (on orbital shaker platforms) and spinner flasks for a total of 3 consecutive passages. Up to a 9-fold increase in cell number was observed over 5 days per passage, with a cumulative fold change up to 600 in 15 days. This expansion would enable literscale bioreactors to be seeded if starting from a 6-well plate suspension culture. Using this process, we were also able to maintain high levels of pluripotency markers (≥95% OCT4) at the end of the expansion, all whilst maintaining a normal karyotype. Additionally, we compared neural induction of hiPSCs by using a dual SMAD inhibition protocol with a commercially available neural induction medium. Since initial aggregate size has an important impact in the commitment of hiPSC into a particular lineage, a previously determined combination of seeding density and agitation rate was successfully used to produce homogeneous populations of hiPSC aggregates with an optimal and narrow distribution of diameters. With StemScale Medium, we were able to obtain up to a 32-fold increase in cell number at the end of a 7-day neural induction protocol within an 80 mL spinner flask. The hiPSC-derived neural progenitors underwent further maturation that stained positive for Tuj1 and were responsive to KCl and Histamine treatments.

The results presented in this work should set the stage for the future generation of a clinically relevant number of human neural progenitors for transplantation and other biomedical applications using controlled, automated and reproducible large-scale bioreactor culture systems.

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Richa Singhania

Weill Cornell Medical Center

Richa Singhania is a cross-disciplinary trained scientist with extensive experience in translational cancer research, and her latest research is based on using stem cells and organoids for cancer modeling and drug discovery. Currently, she is spearheading the Starr Foundation Cerebral Organoid Translational Core at Weill Cornell Medicine, New York which integrates organoid and automation technologies to create personalized solutions for brain cancer patients. Richa received a Masters in Biotechnology and PhD in Molecular Biology from University of Queensland, Australia, and postdoctoral training from academic institutions in the UK (University of Nottingham) and USA (UT Southwestern Medical Center and Weill Cornell Medicine). Richa is passionate about bringing precision medicine to the masses, and making preclinical research successful to ultimately improve the lives of people affected by cancer.


Organoid models of Glioblastoma

Abstract

Discover the latest advances in generating in vitro 3D models of human glioblastoma (GBM). Dr. Singhania will cover how these organoid-based models for the first time in decades are allowing us to capture the characteristic phenotypic and molecular features of GBM in a dish, to ask fundamental questions of brain tumor development as well as to test and exploit therapeutic vulnerabilities of this cancer in a way that cannot be done in traditional cell lines and animal models. Dr. Singhania will also talk about how these models can be personalized to help direct individual patient treatments.





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Jeff Millman

Washington University School of Medicine

Dr. Jeffrey Millman received his Ph.D. in Chemical Engineering from the Massachusetts Institute of Technology and completed his postdoctoral training in the laboratory of Dr. Douglas Melton at Harvard University. He was recruited to Washington University School of Medicine as an Assistant Professor in 2015. His current research is focused on synergizing both biomedical engineering and cell biology approaches to use stem cells for the study and treatment of diabetes. His laboratory is supported by the NIH and JDRF, and he has recently received an American Institute of Chemical Engineering 35 Under 35 Award and Chemical and Biomolecular Engineering Distinguish Young Alumni Award from NC State University. His scholarship has been published in rigorously reviewed journals, such as Nature Biotechnology and Science Translational Medicine, and featured in the New York Post, BBC Radio 4, CBS Evening News, and IFLScience. He has 7 issued patents, many of which have been licensed to biotechnology and startup companies. 


Insulin-Producing Islets to Combat Diabetes from Stem Cells

Abstract

Cellular and tissue engineering promises new therapeutic options for people suffering from a wide range of diseases. Differentiation of stem cells is a powerful renewable source of these functional replacement cells and tissues that can be grown in the laboratory. Diabetes is cause by the death or dysfunction of insulin-secreting islets, which are a tissue type found in the pancreas that contain β cells and other endocrine cell types. We have recently developed approaches combining modulating the actin cytoskeleton and signal transduction pathways during differentiation to produce stem cell-derived islets (SC-islets) capable of undergoing glucose-stimulated insulin secretion, their primary function. We have further expanded this approach to make SC-islets from patients with diabetes and used CRISPR-Cas9 to correct their diabetes-causing mutations. Upon transplantation into mice with severe pre-existing diabetes, these SC-islets rapidly restore normoglycemia and can maintain this functional cure for a year. Our hope is that one day this technology can be used to replace unhealthy islets in patients for therapy and provide a better disease-in-a-dish model to discover new drugs to prevent, stop, or reverse diabetes progression.


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Donna Chang

Hope Biosciences

As the President and CEO of Hope Biosciences, Donna Chang is on a mission to revolutionize the field of cell therapy. She has over 12 years of experience in biotechnology business development, including business expansion and strategic partnering. Donna started her career in economic development in life sciences. She later transitioned into industry, focused entirely on cellular therapeutics. Her passion in this subject is the leading motivation to find solutions to the current limitations in cell therapy and deliver approved therapies to the market - quickly. Donna started Hope Biosciences in 2016 with a goal to develop and deliver adult stem cell therapeutics that are safe, effective and affordable. Hope Bio’s groundbreaking patented core cell culture technology has been utilized in over 12 groundbreaking clinical trials in the United States. The company anticipates to bring these therapies to market in the very near future. Donna graduated from the University of Toronto with a degree in Bioethics and Human Biology. She received her Masters (M.S.) in Biotechnology with a concentration in Enterprise Development from Johns Hopkins University. 


Optimization and Standardization in Stem Cell Manufacturing for the Fight Against COVID-19

Abstract

The pandemic has forced the drug development industry to move at an unprecedented pace. Cellular therapies are finally being looked at as a possibility for immunomodulation and tissue regeneration. However, the existing challenges and limitations in delivering these therapies, are coming to light. Since its inception, Hope Biosciences has been working on solutions to deliver high quality, standardized, fresh mesenchymal stem cells - on -demand. COVID-19 is putting our methods to the ultimate test and gives us a glimpse of what the future looks like.

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Rupa Pike

Thermo Fisher Scientific

As the Head of Technical Operations at Patheon, Rupa oversees the strategic initiatives and solutions for process development (PD) of GMP grade Cell Therapy products. She spearheads technical training and works closely with partners and clients to conduct technology transfer and process optimization activities. Rupa also leads the efforts to build novel Cell Therapy workflows that will advance PD and GMP manufacturing service offerings at Patheon.

Prior to re-joining Thermo Fisher, she worked as the Director of Cell Manufacturing for UW Program for Advanced Cell Therapy. She was part of the leadership team at WiCell Research Institute that pioneered the early human embryonic and induced pluripotent stem cell research under the leadership of Dr. Jamie Thomson. Rupa developed the Stem Cell Training Course which has served over 800 scientists from 32 US states and 20 countries. At Ligand Pharmaceuticals, she was part of the team that designed novel hematopoietic screening assays for erythropoietin and thrombopoietin in partnership with GlaxoSmithKline, a part of the program that led to development of Promacta®. In her past roles, Rupa has successfully managed R&D laboratory operations, Manufacturing Science & Technology (MSAT) operations, technical training,

infrastructure development, customer relations and business development. Rupa holds a Masters degree in Biology from Loyola University and a Doctorate from Harvard University.


Transitioning from Cell and Gene Therapy Discovery and Development to Manufacturing - Challenges and Opportunities

Abstract

Cell and Gene Therapies (CG&T) hold the promise of transforming medicine and human health. The recent commercial success of cell therapy products has generated a tremendous excitement and acceleration of new product development in this arena. Currently, over 400 therapies are in pre-clinical to Phase 3 development, and approximately 1,700 clinical studies are underway globally. As CG&T manufacturing processes evolve to meet regulatory, economic, and patient safety needs, a seamless transition from pre-clinical to GMP manufacturing is critical for their success.

Academic and hospital-based research and development programs play a major role as a source of innovation for new CG&T products. The need for technology transfer and process improvement is foundational as companies acquire licenses for CG&T products from academic centers. The statement that the “process is the product” is particularly true for cell therapies since cells are living pharmaceuticals. The overall success of transitioning the product from development to clinical trials, and ultimately to commercialization, is entirely dependent upon a safe, robust, well-characterized and reproducible process. Scientists can achieve this goal by addressing issues related to raw materials, analytical methods, scalability, personnel training and early incorporation of quality standards. Adopting steps for early process optimization and control will result in manufacturing therapies that meet regulatory requirements and are economically viable at industrial scale.

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Laila Mahmoud Montaser

Menoufia University, Egypt

Laila Mahmoud Montaser MD was born in the City of Alexandria, Egypt, in a family of dynasty doctors, graduated from Faculty of Medicine, Alexandria University and got Diploma degree in Microbiology from Alexandria University, Diploma degree in Clinical Pathology, and MD from Faculty of Medicine, Tanta University. Prof Montaser is a Professor in Clinical Pathology and Hematology; Chair of Stem Cell, Regenerative Medicine, Nano medicine and Tissue Engineering (SRNT) Research Group. She served as the Founder of Clinical Pathology Department Faculty of Medicine, Menoufia University, Egypt, was being the Chair of Department of Clinical Pathology for more than 10 years. She supervised more than 44 M. Sc. theses and 13 Doctorate theses. She has made Faculty of Medicine a hub at Menoufia University. She is uniquely trained and situated and has a philosophy on how to manage research and has written numerous abstracts and research papers on Clinical Pathology, Hematology, and also Stem Cell, Regenerative Medicine, Nano Medicine and Tissue Engineering, her research interest, published in top international journals and Conferences. She has an important intellectual property (IP) on the proposal of a national project to launch an innovative strategy by integration of novel education method (Entrepreneurship Education) in the educational system in Egypt focus on the preparation of youth to meet the requirements of the labor market and develop a plan to implement the education needs associated with the labor market. She is a member of several international and national societies. She is an Honorable Editorial Board Member/Peer Reviewer for many National and International Journals. Montaser’s distinguished career as a Prof, and researcher who had an enormous international impact selected her for three times at 1986, 1998, and 2002 by Council of Menoufia University to Award of “Ideal Doctor” from Egyptian Medical Syndicate and also nominated to two major awards: TWAS prize in Medical Sciences and Nano Science Research Excellence due to her scientific achievements worldwide.

Using Stem Cells in Cartilage Repair and Tissue Engineering

Abstract:

Cartilage repair and renovation stay especially incurable owing to a much weak regenerative prospect of this tissue.  Facing the clinical defy of rebuilding of cartilage disorders, the scope of cartilage engineering has advanced. Until now, the plurality of research into cartilage restoration has been concentrated on articular cartilage due to the considerable predominance of big osteoarthritis in a more and more senility people. Consequently, tissue engineering which targets to originate new and amended tissue and organ subrogate has elicited growing benefit for articular cartilage reform. While stem cells grasp a large possibility for the handling of plentiful lesions and degenerative joint illnesses, various hurdles should be conquered before their medicinal implementation may be recognized. These comprise the evolution of sophisticated processes of advanced techniques to trajectory and evidence implanted stem cells. The fulfillment of nanotechnology to stem cell biology is promising to rubric the challenges of the dud of inoculated cells to contrive to target tissues. Our objective is to construct engineered biomaterial scaffolds to advocate cell culture and create novel cartilage tissue. This manuscript displays existent notions and schemas in cartilage engineering with a confirmation on the employment of stem cells and nanotechnology in the output of biomimetic cartilage reconditioned scaffolds. The domain of Cartilage Tissue Engineering, which is pointed to repair, renovate, and recover damaged or sick cartilage working, has holds exaggerated potential for effective Cartilage-treatment. There is a huge commitment to improve contemporary cartilage medications across attaining a systematically effective tactic for treating cartilage torments. Tissue engineering may be the better mod to make this intent by means of the use of stem cells, novel biologically inspired scaffolds, and emerging nanotechnology.


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Jiayin Yang

Cell Inspire Pharma

Jiayin Yang is currently the Chief Technology Officer of Cell Inspire Therapeutics, a start-up company in Shenzhen, China that aims to develop candidate therapies for neurodegenerative diseases and rare diseases using stem cell-based models. He received his Ph.D. in stem cells and regenerative medicine from the University of Hong Kong. Dr. Yang has 12 years of research experiences in human pluripotent stem cells (PSCs) and genome editing with custom designed nucleases. In combination with these two cutting-edge technologies, Dr. Yang and his team have generated a series of in vitro models with genome engineered induced pluripotent stem cells (iPSCs) for disease models and drug screening.  

Title: iPSC-Based High Throughput Platforms for Screening Novel Therapeutics for Treatment of Alzheimer's Disease and Parkinson’s Disease

Abstract: 

Alzheimer’s disease (AD) and Parkinson’s disease (PD) are complex neurodegenerative diseases that affect millions of people worldwide and are currently no cure. Lacking of suitable cellular models and the inadequate diversity of the existing compound libraries are two hurdles for drug screening for both diseases. To overcome those limitations, we have created novel cellular models based on genetically modified and/or patient-derived human induced pluripotent stem cells (iPSCs). For AD cellular modeling, we site-specifically integrated AD-related genes at the AAVS1 site of a normal iPSC line and created two iPSC lines respectively. Neurons derived from both engineered lines displayed AD phenotypes in vitro, such as significantly elevated Aβ level and p-tau level compared to those of their isogenic parental line.  For PD modeling, we obtained iPSC from a patient with young-onset PD (YOPD) who carried multiple mutations in both PARK2 and HTRA2 genes. Neurons derived from the YOPD iPSC displayed typical PD phenotype and severe neurodegenerative features. In order to create high throughput platforms suitable for drug screening, we have also established standardized processes for production of large quantities of uniform neurons from AD and PD iPSC lines and developed robust human neuron-based assays for drug candidate screening. 

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Soong Poh Loong

Ternion Biosciences Pte. Ltd.

Poh Loong Soong is the co-founder of Ternion Biosciences Pte. Ltd., a Singapore startup biotech company which focus on innovative technologies for accelerated drug development and screening. PL’s expertise in human stem cell derived cardiomyocytes have contributed to the development of a novel platform-assay OptioQUANT, enabling precise high throughput fluorescence based readouts from physiologically matured cells at single cells resolution. As a result, Ternion was awarded the ESG-TECs POV grant for this development. Having direct involvement in the development of the cGMP grade human stem cell lines in ES Cell International previously, PL has deep insights and expertise in human stem cell lines generation, genetic engineering and pharmaco-disease modelling. PL obtained his Dr. rer. nat. from the Dept of Pharmacology and Toxicology, UMG at the University of Göttingen in AG. Prof. Zimmermann’s Lab and was responsible for establishing the hSC culture capabilities in the department. As a senior scientist, PL has led efforts to advance hSC technologies particularly in regenerative medicine using tissue engineering approaches and holds a patent for the generation and fabrication of a novel biomimetic BioVAD for provision of cardiac restraint. He is currently based in Singapore in the iHealthTech department at the Yong Loo Lin School of Medicine of the National University of Singapore.

Human stem cell derived cardiomyocytes for drug screening and tissue engineering

Abstract:

Human stem cell derived cardiomyocytes and engineered heart muscle offers great potential to model human disease, drug discovery and safety screening. We routinely culture human induced pluripotent stem cells in animal component free medium followed by mesodermal induction, direct differentiation and cardiac specification either as cell monolayers or small aggregates.

At Ternion, we culture these cardiomyocytes on our custom built OptioQUANTTM platform and perform characterizations to subtype into pacemaker, atrial and ventricular cells. Together with our collaborators, the platform was validated in the article published in Stem Cells Research & Therapy.

For drug screening purposes, we first acquired baseline membrane action potentials of the cardiac subtypes, followed by exposures of drug/compounds at pharmacological concentrations. OptioQUANTTM acquires sub-second fluorescence signal changes with single cells resolution, tenabling precise reports of subtle drug induced changes in membrane action potentials from large numbers of human CMs simultaneously. This allows us to screen compounds with precision and model the kinetics of the drug Mechanism of Action (MoA) in an unbiased manner.

To provide a more physiological model of the heart tissue, we exploited the regenerative potential of hSC-CMs to generate 3D constructs engineered heart muscles in. These engineered heart muscles can then be used for drug screening, or as an epicardial cardiac patch to render contractility in a myocardial infarcted (MI) heart.

Taken together, human stem cell derived cells provide a viable alternative to animal based disease modeling, leading to safer drug development and holds great therapeutic potential for human disease.

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Kwang-Soo Kim

Molecular Neurobiology Laboratory, McLean Hospital/Harvard Medical School

Kwang-Soo Kim is Professor and Director of the molecular neurobiology laboratory at McLean Hospital/Harvard Medical School. Based on his >30 years’ experiences to investigate the transcriptional regulatory cascade of midbrain dopamine neuronal system, he has been focusing on translating his research to novel therapeutic development of Parkinson’s disease (PD). In particular, since the loss of a specific cell type, (i.e., A9 dopamine neuron in the substantia nigra) directly causes the major motor symptoms of PD, his team is focused on human pluripotent stem cell (hiPSC) research and hiPSC-based personalized cell therapy. 

Toward this long-term goal, he is working on establishing an efficient platform to establish the safety and efficacy of hiPSC-based cell therapy for PD. In this seminar, Dr. Kim will share his experience of the first PD patient treated with his own skin cells.

Personalized cell therapy for Parkinson’s disease

Abstract

Parkinson's disease (PD) is a neurodegenerative disorder associated with loss of midbrain dopamine (mDA) neurons in the substantia nigra, rendering cell transplantation a promising therapeutic  strategy. Toward human induced pluripotent stem cell (hiPSC)-based autologous cell therapy for PD, we established a platform of core techniques for the production of mDA progenitors as a safe and effective therapeutic product, including an improved reprogramming method, a novel “spotting”-based in vitro  differentiation methodology, a chemical method that safely eliminates undifferentiated hiPSCs, and an improved neurosurgical device. Using this platform, we extensively studied and confirmed the in vivo safety and efficacy in rodent models of PD. Then, we performed the implantation of patient derived mDA progenitor cells, differentiated in vitro from autologous iPSCs, in a patient with idiopathic PD. The patient-specific progenitor cells were produced under good manufacturing practice conditions, characterized by phenotypic properties for mDA neurons, and implanted into the putamen bilaterally 6 months apart. No immunosuppression was used. PET scans using 18F-DOPA suggested graft survival. Clinical measures of Parkinson’s symptoms after surgery have improved at 18-24 months after implantation together with the patient’s quality of life.

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Michael Akenhead

Thermo Fisher Scientific

Michael Akenhead is currently a Scientist III in the Biosciences Division at Thermo Fisher Scientific.

In 2016, Michael received a PhD in Biomedical Engineering from the University of Kentucky. After completion of his PhD, he was hired to work at Thermo Fisher Scientific.

His primary research involves working with stem cells. Specifically, Michael is exploring the ability to culture stem cells in suspension using new media formulations. The results of this research have led to the development of the GibcoTM StemScaleTM PSC Suspension Medium prototype.

Addressing the Needs of 3D Suspension Culture Systems Through the Optimization of a New PSC Medium.

Abstract:

As the use of pluripotent stem cells (PSCs) in therapeutic and screening applications continues to expand, a key bottleneck is the efficient generation of high-quality PSCs. PSC suspension culture offers key advantages for scale up over adherent culture systems: overall cost; reduced footprint and hands on time; and compatibility with closed systems. In addition, expansion of PSCs in suspension culture consumes less media (and plasticware) than the same number of cells grown in adherent systems. Together, these features make suspension culture an attractive path for cost-effective and rapid generation of large quantities of cells required for downstream applications. 

Although suspension culture offers significant benefits, wider adoption has been limited due to a number of barriers. In particular, there is a lack of effective culture systems and protocols which support PSC suspension culture and scale up. To help address this gap, we have recently developed a new suspension culture media system – Gibco™ StemScale™ PSC Suspension Medium, which promotes the efficient self-aggregation of singularized PSCs into spheroids, without the need for microcarriers. Spheroids grown in this media maintain robust cell expansion (≥ 8 fold). In addition, the viability and pluripotency remain high (≥ 90% of cells) for spheroids cultured over consecutive passages. We also demonstrate our new system is compatible with a variety of culture vessels (from well plates to bioreactors) through the optimization of straightforward parameters. We have found PSCs can be maintained in small volume suspension cultures or readily scaled up into large volume culture vessels. 

The StemScale PSC Suspension Medium maintenance, feeding and passaging protocols are designed to be simple processes that minimize the risks of cell loss and contamination and that are also scalable and amendable to automation. During routine culture, singularized cells can be replated into adherent cultures, cryopreserved, and/or reseeded back into suspension conditions for continued expansion. 

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Naomi Okugawa

Biogen, Cambridge, MA

Naomi Okugawa PhD is a Scientist at Biogen, Cambridge, MA.  She obtained her PhD in Japan, then performed postdoctoral training at Massachusetts General Hospital and Novartis.  She worked and learned human iPSC at Harvard Stem Cell Institute.  Her focus is establishing better iPSC models to understand neurological disorders for drug discovery, and her group has been supporting various disease research groups at Biogen by providing iPSC-derived neurons and glial cells.

Characterization of human iPSC-derived neurons by transcriptomics and proteomics for drug discovery

Abstract:

To build a credible chain of translation in which the in vitro model captures the molecular mechanism of a clinical condition, it is necessary to have physiologically relevant human cell models that can be produced in a robust and reproducible manner. In the past, this has been particularly difficult to achieve for neurons and other cell types of the central nervous system. It has been shown that overexpression of a single transcription factor, neurogenin 2 (NGN2), can drive differentiation of undifferentiated induced pluripotent stem cells (iPSC) into highly pure populations of cortical excitatory glutamatergic neurons.  We adapted this method to produce iPSC-derived neurons with minimal batch-to-batch variability at a scale sufficient to support drug discovery programs in the biomedical industry. 

To that end, a doxycycline-inducible expression cassette was genetically integrated into the AAVS1 locus of a healthy control iPSC line, 7189L. After expansion of 7189L iPSC, differentiation was started by overexpressing NGN2 for three days before cells were cryopreserved. Five batches of partially differentiated neurons were recovered, matured, and characterized for consistency in gene expression (RNA-seq), protein expression (nLC-MS/MS) and electrophysiological function by microelectrode array (MEA) technology in a time course experiment over five weeks in culture. Both proteomics and RNA-seq analyses showed progressive neuronal maturation in a manner that was highly consistent between production batches.  More than 11,000 proteins were identified in the proteomics study, spanning an abundance range of over seven orders of magnitude. Proteomics data showed an upregulation of proteins associated with neuronal function such as synapse formation and neurotransmitter production throughout the time course.  In lines with these observations, time-dependent upregulation of axonal guidance and synaptogenesis were most enriched gene sets in the RNA-seq data.  MEA recordings demonstrated robust network activity of the neurons after 28 days of maturation and confirmed the presence of AMPA dependent network bursting, consistent with a pure population of glutaminergic excitatory neurons. In summary, we have established a robust, reproducible and scalable protocol to generate human cortical neurons which were fully characterized by proteomics and transcriptomics for disease modeling and assay development.  

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Dirk Walther

Biogen, Cambridge, MA

Dirk Walther obtained his PhD in the group of Matthias Mann at the Max-Planck Institute of Biochemistry in Germany and has been working as a staff scientist in the Chemical Biology and Proteomics group at Biogen in Cambridge, Massachusetts since 2014. He applies quantitative proteomics workflows to support early through late stage drug discovery programs, performing experiments to discover and validate drug targets, and determine the mechanism of action of drug candidates.

Characterization of human iPSC-derived neurons by transcriptomics and proteomics for drug discovery

Abstract:

To build a credible chain of translation in which the in vitro model captures the molecular mechanism of a clinical condition, it is necessary to have physiologically relevant human cell models that can be produced in a robust and reproducible manner. In the past, this has been particularly difficult to achieve for neurons and other cell types of the central nervous system. It has been shown that overexpression of a single transcription factor, neurogenin 2 (NGN2), can drive differentiation of undifferentiated induced pluripotent stem cells (iPSC) into highly pure populations of cortical excitatory glutamatergic neurons.  We adapted this method to produce iPSC-derived neurons with minimal batch-to-batch variability at a scale sufficient to support drug discovery programs in the biomedical industry.  

To that end, a doxycycline-inducible expression cassette was genetically integrated into the AAVS1 locus of a healthy control iPSC line, 7189L. After expansion of 7189L iPSC, differentiation was started by overexpressing NGN2 for three days before cells were cryopreserved. Five batches of partially differentiated neurons were recovered, matured, and characterized for consistency in gene expression (RNA-seq), protein expression (nLC-MS/MS) and electrophysiological function by microelectrode array (MEA) technology in a time course experiment over five weeks in culture. Both proteomics and RNA-seq analyses showed progressive neuronal maturation in a manner that was highly consistent between production batches.  More than 11,000 proteins were identified in the proteomics study, spanning an abundance range of over seven orders of magnitude. Proteomics data showed an upregulation of proteins associated with neuronal function such as synapse formation and neurotransmitter production throughout the time course.  In lines with these observations, time-dependent upregulation of axonal guidance and synaptogenesis were most enriched gene sets in the RNA-seq data.  MEA recordings demonstrated robust network activity of the neurons after 28 days of maturation and confirmed the presence of AMPA dependent network bursting, consistent with a pure population of glutaminergic excitatory neurons. In summary, we have established a robust, reproducible and scalable protocol to generate human cortical neurons which were fully characterized by proteomics and transcriptomics for disease modeling and assay development.  

 

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Thor Theunissen

Washington University School of Medicine

Thor Theunissen grew up in The Netherlands and received his A.B. in Biology from Harvard in 2007. He became interested in stem cells and developmental biology during his undergraduate work in the laboratories of Christine Mummery (Hubrecht Institute) and Stuart Orkin (Harvard Medical School). He completed his graduate studies in José Silva’s laboratory in the Wellcome Trust Center for Stem Cell Research and Department of Biochemistry at the University of Cambridge in 2011. His doctoral thesis focused on the role of the homeodomain transcription factor Nanog in epigenetic reprogramming. As a Sir Henry Wellcome Postdoctoral Fellow in Rudolf Jaenisch’s laboratory at the Whitehead Institute/MIT, Thor developed methods to isolate naïve human pluripotent stem cells. He was appointed Assistant Professor in the Department of Developmental Biology and Center of Regenerative Medicine at Washington University School of Medicine in 2017. He is a recipient of the NIH Director’s New Innovator Award (DP2), the Edward Mallinckrodt Jr New Investigator Award and the Shipley Foundation’s Program for Innovation in Stem Cell Science Award.

Understanding human pluripotent states and their applications

Abstract:

The past decade has seen significant interest in the isolation of pluripotent stem cells corresponding to various stages of mammalian embryonic development. Two distinct and well-defined pluripotent states can be derived from mouse embryos: "naïve" pluripotent cells with properties of pre-implantation epiblast and "primed" pluripotent cells, resembling post-implantation epiblast. Prompted by the successful interconversion between these two stem cell states in the mouse system, several groups have devised strategies for inducing a naïve state of pluripotency in human pluripotent stem cells. Here, I will review recent insights into the naïve state of human pluripotency and its applications in biomedical research. The isolation of naïve human pluripotent stem cells offers a window into early developmental mechanisms that cannot be adequately modeled in primed cells, such as X chromosome reactivation and the regulation of hominid-specific transposable elements. Furthermore, recent work from our lab and others indicates that naïve hPSCs have a unique potential to differentiate into extraembryonic lineages, including trophoblast and primitive endoderm. These advances provide a platform to model cell fate decisions in the early human embryo and elucidate the origins of common pathologies afflicting placental development.

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Zhexing Wen

Department of Psychiatry and Behavioral Sciences at Emory University

Dr. Zhexing Wen received his PhD training at the Rutgers University in 2008. In 2009, he joined Drs. Hongjun Song and Guo-li Ming’s laboratories at Johns Hopkins University as a postdoctoral fellow. In 2016, Dr. Wen joined the Department of Psychiatry and Behavioral Sciences at Emory University as an Assistant Professor. The research of Dr. Wen’s laboratory centers on understanding the molecular mechanisms underlying neuropsychiatric disorders. In particular, he is interested in using patient-derived induced-pluripotent stem cells (iPSCs) to elucidate the biological functions of causal/risk genetic and environmental factors in neuropsychiatric disorders, identify pathological developmental processes that may contribute to the etiology of these complex diseases, and translate such knowledge into therapeutic targets for developing novel treatments.

Molecular dynamics associated with development of human GABAergic interneurons

Abstract:

GABAergic interneurons (GINs) are a heterogeneous population of inhibitory neurons that collectively contribute to the maintenance of normal neuronal excitability and network activity. Several homeobox and basic helix-loop-helix transcription factors are known to contribute to neural patterning and early GIN fate specification, including DLX1/DLX2, NKX2.1, ASCL1, and MASH1. However, much less is known about the networks of transcription factors and genetic elements that contribute to mature GIN function. In particular, there is a knowledge gap in the epigenomic dynamics of developing GINs, which may direct interneuron-specific expression patterns. Identification of these regulatory components may provide new insight into the pathways underlying proper GIN activity, while also denoting potential therapeutic targets for GIN-associated disorders, such as schizophrenia and epilepsy. Here, we differentiated human induced pluripotent stem cells (iPSCs) derived from two healthy male controls into GINs with 81-85% efficiency. To examine temporal changes in gene expression and chromatin accessibility, cells were collected at three time points for RNA-seq and ATAC-seq analysis: neural progenitor cells (NPCs) at 22 days post-differentiation (D22), then GINs at D50 and D78. By comparing differentially accessible regions (DARs) of chromatin that were shared between the two iPSC lines, we identified 13,221 genomic regions that correlated with temporal changes in gene expression unique to mature GINs. We also classified several transcription factors (TFs) that were increasingly enriched at DARs during differentiation, indicating regulatory networks that may underlie GIN function. Furthermore, we identified several genes that may be especially relevant to mature GIN function in schizophrenia patients. Collectively, these data represent a comprehensive analysis of transcriptomic and epigenomic changes that occur during GIN development.

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David Kuninger

Thermo Fisher Scientific

Director, R&D and Lead of Cell Models within the Cell Biology Business of Thermo Fisher Scientific.

Seasoned Biotechnology professional with broad experience driving innovation, product development and commercialization. Currently lead multiple R&D teams supporting Stem Cell Culture and Differentiation, Neurobiology, Primary Cells & ADME/Tox, 3D Biology and Cell Therapy applications incorporating these cell types. In the last 5 years his group has been responsible for the launch of over 40 new culture system products spanning a diverse array of biological applications.

 Differentiation of iPSCs in 3D: Leveraging Suspension Cultures for Scale and Efficiency

Abstract:

As the use of pluripotent stem cells (PSCs) in therapeutic and screening applications continues to expand, a key bottleneck is in the efficient generation high-quality PSCs. Three-dimensional (3D) suspension cultures offer several key advantages for scale up over two-dimensional (2D) adherent culture including overall cost, reduced footprint and hands on time, as well as compatibility with closed systems and reduced risk of contamination.   In addition, expansion of PSCs in 3D cultures consumes less media (and plasticware) than the same number of cells grown in 2D cultures. Together, these features make 3D culture an attractive path for cost-effectively and rapidly generating large quantities of cells required for certain downstream applications. We have recently developed a new 3D suspension culture medium – Gibco™ StemScale Medium, which was designed to promote the efficient self-aggregation of singularized PSCS into spheroids, without the need for microcarriers, while maintaining robust growth per culture level.  Both viability and pluripotency remain high for spheroids cultured in this medium over consecutive passages.   Here we show these expanded iPSC spheroids can be taken directly into various directed differentiation protocols while maintained in 3D suspension culture.  Our results demonstrate highly efficient induction into all three germ lines and subsequent downstream generation of cardiomyocytes and neurons as specific examples.  Strategies and approaches for adapting protocols designed for adherent culture for use in 3D suspension applications as well as potential benefits of “toggling”  between 2D and 3D culture are discussed.

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Yanhong Shi

Beckman Research Institute of City of Hope

Yanhong Shi is Herbert Horvitz Professor in Neuroscience and Director of Division of Stem Cell Biology Research at Beckman Research Institute of City of Hope. She earned her Ph.D. degree from Northwestern University. Upon graduation, she went to the Salk Institute for postdoctoral training, where she studied nuclear receptors in neural stem cells and neurogenesis. Her lab at Beckman Research Institute of City of Hope focuses on human iPSC-based disease modeling, drug discovery, and cell therapy development for neurological diseases. 

Human IPSC-Based Disease Modeling and Therapeutic Development

Abstract:

The iPSC technology has provided great hope for developing cell therapies, in addition to serving as a platform for disease modeling and drug discovery. We have used the human iPSC platform to model neurological disorders and develop cell therapies for neurological diseases. For example, we have established stem cell therapy candidates for Canavan disease (CD), a devastating neurological disease that has neither a cure nor a standard treatment. We established a cGMP-compatible manufacturing process for human iPSC derivation and subsequent differentiation and genetic modification. We further tested the human iPSC-derived cellular products in a Canavan disease mouse model and demonstrated robust efficacy and preliminary safety of the cellular products. This study could provide an effective therapeutic approach, and eventually lead to a cure for Canavan disease, and other related neurological diseases.


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Johan E. Morales-Sánchez

Biotechnology Research Center, Costa Rica Institute of Technology

Johan Morales-Sánchez is a biotechnology engineer graduated in 2014 from the Costa Rica Institute of Tecnology (TEC), who currently works as a researcher in the Biotechnology Research Center from TEC. During the last years, he has taught pre-grad students in this university about Biology and Cell Culture, and has also been focused in tissue engineering and laboratory animal research.

Stem cell therapies for skin regeneration in animal models, Costa Rica experiences

 Abstract:

The Tissue Engineering Laboratory (LAINTEC) of the Costa Rican Institute of Technology (TEC), started activities in 2005 aiming to develop cell therapies focused on skin regeneration. Throughout the last fifteen years, this laboratory has grown and the research has diversified, so now it has more than a dozen of multidisciplinary projects focused on regenerative skin therapies, development of three-dimensional models of skin, muscle and bone, evaluation of new biomaterials, among other research lines. One of the most important areas is the development of skin regeneration therapies. As part of the experience, we have worked with two approaches using different types of stem cells: epidermal stem cell (ESC) and adipose derived stem / stromal cell (ASC). First, ESC were isolated, expanded and characterized analyzing cytokeratines 10, 14 and 15 expression. To evaluate the regeneration potential of ESCs, a controlled full thickness skin wound was performed in the interscapular area in an adult rat model and a plasma-based matrix (PBM) with fibroblasts and keratinocytes applied, compared with a PBM with fibroblasts or PBM alone as control. On the other hand ADSC were isolated and the identificated using CD73, CD90, CD105 and CD45 markers. Also, the differentiation potential was analyzed. To evaluate the regeneration using ASC, 5 six-week-old BalbC mice per group were subcutaneously treated with: ASC in saline solution, ASC seeded on a natural scaffold and scaffold with saline solution. A positive and negative control was also performed. After two weeks of treatment, at a macroscopic level, both treatments showed no significant differences in the regeneration rate. However, the ASC treated group, showed more vascularization and a better collagen fibers organization compared with the control groups. Eventhough is necessary more preclinical testing and evidence, these experiments present the first steps taken in Costa Rica towards cellular therapy development as well as strengthening research in regenerative medicine and tissue engineering.

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Carl Dargitz

Thermo Fisher Scientific

Carl Dargitz currently works at Thermo Fisher Scientific as a Scientist III in the Cell & Gene therapy workflows group.  During his four years at Thermo Fisher Scientific he has worked to develop characterization assays for PSCs and immune cell types and on cell therapy workflows and instrumentation.  He received his Master’s degree from California Polytechnic State University, San Luis Obispo in Biomedical Engineering with a specialization in stem cell research in 2013.  Previously, he worked in Dr. Inder Verma’s lab at the Salk Institute differentiating iPSCs into lung airway epithelium to study Cystic Fibrosis, and at the stem cell core facility at Salk managing the core’s reprogramming and characterization services.   

Rotea: A Versatile Cell Therapy Workflow Instrument for Cell Isolation, Washing, and Concentration

Abstract:

Bringing cell and gene therapy treatments to patients has many challenges to overcome, however, none may be greater than the efficient and consistent processing and delivery of these products.  Rotea’s counterflow centrifugation technology enables cell separation, washing, and concentration of many cells types and can be easily programmed to serve many additional workflows.  The single use kit and instrument are able to capture and sustain up to 5 billion cells in a stable, fluidized bed with flexible inputs and output volumes as low as 5 mL. Protocols can be designed to wash and concentrate virtually any cell type, including various types of stem cells by adjusting the centrifuge and pump rate.