We’re Heading to ISSCR Next Week – Are you? Don’t miss these great talks and activities
June 16, 2016
ISSCR’s Annual Meeting begins June 22nd in San Francisco. The ISSCR Annual Meeting is “a place where breakthrough science, global collaboration, and researchers across the breath of the field come together. The ISSCR Annual Meeting offers the best opportunity to collaborate with leaders and peers from around the world to fully realize the promise of our field in therapies, treatments, and clinical advances.” Cell Culture Dish will be attending and blogging from the event. Please don’t miss our ISSCR related blogs over the upcoming months.
If you are attending, don’t miss these exciting talks and activities!
Interesting Talks in Each Track. Here are some examples:
Presidential Symposium – Stem Cells and Cancer Tissue Growth and Morphogenesis Cellular Plasticity and Reprogramming Gene Networks and Epigenetics Gene Therapy and Stem Cells Disease Modeling Using Stem Cells Cell Therapy in Clinical Trials
Concurrent Session I
Neural Stem Cells Paradigms from Model Organisms Mechanisms of Pluripotency Disease Modeling I Stem Cell Plasticity Muscle Stem Cells Cardiac Development and Regeneration
Concurrent Session II
Neural Disease Stem Cells in Organ Development and Maintenance Mechanisms of Asymmetric Cellular Division Nutritional Effects on Stem Cells Stem Cell Niches Stem Cells and Solid Cancers Development and Regeneration in Endodermal Derivatives
Concurrent Session III
Neurodevelopment and Regeneration Germline Stem Cells Stem Cell Aging Direct Cellular Reprogramming Skeletal Stem Cells Hematopoiesis Epithelial Development and Regeneration
Concurrent Session IV
Road to the Clinic Tissue Engineering Stem Cell Metabolism Regulatory Networks in Differentiation and Disease Disease Modeling II Leukemia and Stem Cells Technology Frontiers
Cell Culture Dish Sponsor Activities:
BD Biosciences – Booth 1625
Biological Perspectives in Multicolor Flow Analysis: Moving to 50-color Flow CytometryThursday, 23rd June, 11:30am – 12:30pmBob Balderas, VP Biological Sciences, BD Biosciences
The advent of human genome sequencing and analysis have greatly enhanced our understanding of the underlying biochemistry of complex biological systems. More recently, significant advances in molecular and cellular analysis have continued to deepen our scientific insights in many different biological fields, including immune system function in normal and disease states. The development of the BD FACSymphony™ flow cytometer, with capabilities to analyze up to 50 parameters, now provides a powerful approach to unravel highly complex molecular structure and function relationships in genomics, proteomics, systems immunology, and biology. This research approach holds the promise of increasing our knowledge of cancer biology, accelerating vaccine development, and enhancing the drug discovery process.
As a result of the availability of new BD Horizon Brilliant™ polymer fluorochromes, a study of receptor density and expression, and the launch of the BD FACSymphony high parameter cell analyzer, this year we demonstrated practical 27-color flow cytometry. In this presentation, we will discuss a systematic strategy for successful panel design for high parameter multicolor assays.
Biological Industries– Booth 1419
An Innovative Xeno-Free Culture Medium for the Expansion of Cell Therapy-Compliant Human Endothelial CellsFriday, June 24, 11:30 AM-12:30 PM Room 2002Dr. Sharon Daniliuc, R&D, Biological Industries
Human endothelial cells (hEC) form a single cell layer that lines the interior surface of blood and lymphatic vessels, termed endothelium. Creating a functional vascular network in vitro is crucial for preventing ischemia in engineered tissues. Consequently, EC have become a key element in tissue engineering and cell-based therapy to improve graft implantation. Currently, there are no efficient xeno-free (XF) medium for the growth and expansion of hEC. A defined XF culture system optimized for hEC expansion would greatly facilitate development of a robust, clinically accepted culture process for quality-assured cells.
The present study evaluated a novel XF culture system comprising of hEC medium (EndoGo™ XF Medium) and auxiliary solutions for attachment, dissociation, and cryopreservation of hEC. Results show that the hEC culture system efficiently supports long-term expansion of hEC from large vessels (arteries and veins, e.g. HUVEC) and small vessels (lymph and blood). Expanded cells maintain EC features: typical cobblestone-like cell morphology, phenotypic surface marker phenotype, gene expression profile, and capillary-like structure formation.
A Novel Set of Serum-Free, Xeno-Free Differentiation Media for Adipogenesis, Osteogenesis and Chondrogenesis of Human Mesenchymal Stem Cells from Various Tissue Sources
The poster presents the development of novel serum-free, xeno-free media and supplements that efficiently differentiate hMSCs from various sources into adipocytes, osteoblasts, and chondrocytes. Molecular characterization and functional assays were used to evaluate the quality and purity of the differentiated cells.
A Xeno-Free Culture System for hMSC from Various Sources Suitable for Initial Isolation and Expansion Toward Clinical Applications
The poster presents an evaluation of a novel xeno-free culture system, comprising MSC NutriStem® XF culture medium and all the required auxiliary solutions for the attachment, dissociation, and cryopreservation of the cells. The system was evaluated for initial isolation of hMSC from various sources, and for long-term culturing under SF, XF culture conditions suited for clinical applications.
A Xeno-Free CultureSystem for Human Microvascular Endothelial Cells from Various Sources
The poster presents an evaluation of a novel xeno-free culture system comprising microvascular EC XF medium (EndoGo™ XF Medium) and auxiliary solutions for attachment, dissociation, and cryopreservation of hMVEC. The system was evaluated for long-term culturing of EC from various sources under XF culture conditions.
For more information about Biological Industries’ Activities at ISSCR, please click here
Corning – Booth 1619
Stem Cell Culture, Differentiation and Scale-up – Novel Technologies Enabling Research and Cell Processing ApplicationsFriday, June 24th from 11:30am-12:30pm – Level 2, Rooms 2014/2016/2018
A critical component for basic and clinical research are stem cells that maintain an undifferentiated phenotype during expansion followed by differentiation to specific cell types. In this session we will discuss robust, easy to use, reproducible, scalable systems, including feeder-free and novel animal-free platforms for pluripotent and adult stem cell expansion and differentiation, including Matrigel Matrix, pre-coated recombinant Laminin521, Fibronectin/ Collagen-I / Vitronectin peptide mimetic cultureware, hMSC expansion media, scalable vessel platforms and closed systems.
For more information about Corning’s Activities at ISSCR, please click here
Irvine Scientific – Booth 1509
Optimizing Culture Medium That Maintains Mouse Hematopoietic Stem Cells
Thursday, 23rd June 11:30am – 12:30pm – Moscone West, Level 2, room 2002Dr. Hideyuki Oguro, Department of Pediatrics, University of Texas Southwestern Medical Center
Hematopoietic stem cell expansion in culture is a critical yet challenging goal to achieve to facilitate Gene Therapy and increase the transplantation safety and efficacy. In this workshop, we will share the evaluation results of culturing 20 isolated, CD150+CD48-/lowCD34-/lowLineage-Sca-1+c-kit+ highly enriched HSCs from young-adult mouse bone marrow in prototypes media comparing to the commonly used SF-O3, supplemented with SCF and TPO. Data from colony assay and transplantation studies demonstrated a simplified, ready-to-use medium that could allow us to continue fine tuning the culture components to further improve HSC maintenance ex vivo.
Impacts of Undefined Components for Primary Cell CulturesJessie H.-T. Ni, Ph.D. Chief Scientific Officer, Irvine Scientific
Natures of stem/progenitor cells generated from ex vivo cultures are critically dependent on the quality of supplemented serum or serum-replacement components, e.g. cytokines or lipids. We will demonstrate through historical data and case studies, how these “undefined” components impact the culture performance of major adult stem cells and immune cells. Potential solutions will be recommended that could allow researchers to best achieve consistent and desired ex vivo expansions of targeted stem/primary cells under less-defined culture conditions.
From Research to GMP Manufacturing: Covering All the BasesFriday June 24th, 8:00-8:30
Julie Murrell, Ph.D., Head of Cell Therapy Bioprocessing, MilliporeSigma Ethan Patterson, Ph.D., Senior R&D Scientist, MilliporeSigma
When making the translation from research to the clinic, demonstrating efficacy is only the first step to a successful product. Long-term commercial viability requires high quality materials that offer a safe and sustainable supply coupled with early process scalability. The Cell Design Studio specializes in genome engineering, enabling alterations in patient-derived iPSCs that introduce or correct disease-related mutations in order to make ideal isogenic cell models. Here we will provide examples for stem cell engineering, focusing on knockouts, targeted integration, and SNP introduction/correction along with the validation process and downstream differentiation options. This provides researchers with a useful tool for high content analysis, high throughput screening, target validation, and other cell-based assays. We will also present a model for translating small scale Cell Therapy processes to a robust manufacturing process including media, reagents, bioreactor expansion, and harvest. A well thought out strategy for GMP processing requirements from phase I trials to commercialization is essential in successfully translating a potential therapy to a commercial product.
StemBioSys – Booth 1513
StemBioSys BM-HPME®: A Novel 3-Dimensional Microenvironment to Enhance Mesenchymal Stem Cell ExpansionSy Griffey, StemBioSys, COO & Bob Hutchens, StemBioSys, CEO
StemBioSys, Inc. is a privately-held biomedical company focused on enhancing the growth and delivery of adult stem cells for research, therapeutic or drug discovery applications. Its patented and proprietary technology platforms overcome key obstacles to creating clinically useful stem cell therapies. The centerpiece of these enabling technologies is the HPME® (High Performance Micro Environment) platform. This cell derived 3-dimentional microenvironment allows a variety of stem cells to replicate more rapidly, maintain a small cell size and express markers indicative of retained stem cell potency beyond that seen with traditional tissue culture substrates. This technology is positioned to transform and improve the methods and cost of growing and delivering various stem cell populations for applications in research, therapeutic and drug discovery.
#W3033 Stromal Cell-Derived Extracellular Matrix Stimulates Mesenchymal Stem Cell Proliferation In Vitro And Bone Formation in Rat Femur Segmental Defects –Wednesday, June 22, 2016, from 6:30 PM to 7:30 PM within the ‘Tissue Engineering’ section at Poster Session I Odd
Sy Griffey, PhD #F1038 Stromal Cell-Derived Extracellular Matrix Promotes Cord Blood-Derived Mesenchymal Stem Cell Proliferation While Restricting Neural Lineage Differentiation –Friday, June 24, 2016, from 7:00 PM to 8:00 PM within the ‘Mesenchymal Cell Lineage Analysis’ section at Poster Session III Even
Sy Griffey, PhD #T3034 Rescuing the Regenerative Capacity of Human Mesenchymal Stem Cell Populations from Elderly Donors –Thursday, June 23, 2016, from 7:00 PM to 8:00 PM within the ‘Tissue Engineering’ section at Poster Session II Even
Travis Jackson Block, UTHSCSA#T3019 To Each it’s Own: Tissue Specific Extracellular Matrix Provides an Optimal Niche for Studying Mesenchymal Stem Cell Derived from Bone Marrow and Adipose Tissues –Thursday, June 23, 2016, from 6:00 PM to 7:00 PM within the ‘Technologies for Stem Cell Research’ section at Poster Session II Odd
Milos Marinkovic, UTHSCSA#F4002 Proteomic Characterization of Stem Cell-Derived Extracellular Matrices –Friday June 24th from 7 to 8 PM
Heloise Ragelle, PhD, MIT
Stemcell Technologies – Booth 1633
BrainPhys™ Neuronal Medium Supports Neurophysiological Activities of Human Pluripotent Stem Cell-Derived Neurons
Thursday June 23, 8am – 8:30am | Level 2, Room 2009/2011Cedric Bardy, Research Associate, Salk Institute for Biological Studies
Neurons derived from human pluripotent stem cells (hPSCs) are useful in vitro models for the study of neurological disease and development. To increase the relevance of these studies, it is important that neurons are cultured in a medium that closely resembles the normal physiological conditions of the brain. Traditional media, such as Neurobasal and DMEM/F-12, were designed for neuronal survival, but do not support neurophysiological activities. To resolve this problem, we have developed a new basal medium formulation, BrainPhys™. When supplemented with the appropriate growth factors, BrainPhys™ sustains cell survival and optimal activity of neurons in vitro. Furthermore, electrophysiological recordings and calcium imaging studies showed that the neuronal activities suppressed in Neurobasal and DMEM/F-12-based cultures could be rescued when the media were replaced with a BrainPhys™-based medium. Together, these data show that BrainPhys™ is a novel basal medium that efficiently supports the growth of neurons under conditions that are truly physiologically compatible to that of the central nervous system.
STEMdiff™ Kits for Robust and Efficient Differentiation of Human Pluripotent Stem Cells to Multiple Cell Types
Friday June 24, 8am – 8:30am | Level 2, Room 2009/2011Melanie Kardel, Scientist, STEMCELL Technologies
Human pluripotent stem cells (hPSCs) have many applications in developmental biology studies, disease modeling, drug screening and regenerative medicine. Advancement in each of these areas requires robust and efficient differentiation methods to direct hPSCs to specific cell types of interest. STEMCELL Technologies’ STEMdiff™ kits are standardized reagents that are rigorously optimized for robust differentiation to specific cell types derived from the three germ layers: ectoderm, mesoderm and endoderm. In this showcase presentation, we will highlight the development of our most recent STEMdiff™ products. These include the STEMdiff™ Trilineage Differentiation Kit, a simple culture assay for rapid assessment of pluripotency, as well as new kits for directed differentiation to several mesodermal lineages: STEMdiff™ Mesoderm Induction Medium, STEMdiff™ Hematopoietic Kit and STEMdiff™ Mesenchymal Progenitor Kit. Finally, we will discuss the expansion of our endodermal-related STEMdiff™ system to include the STEMdiff™ Pancreatic Progenitor Kit. In summary, this tutorial will highlight STEMCELL Technologies’ expanding differentiation product portfolio for hPSC research.
Expansion of Human Pluripotent Stem Cells as Aggregates in mTeSR™3D Suspension Cultures
Friday June 24, 11:30am-12pm | Level 2, Room 2009/2011Eric Jervis, Principal Scientist, STEMCELL Technologies
Carrier-free 3D suspension culture enables the convenient scale-up of human pluripotent stem cell (hPSC) production. STEMCELL Technologies has developed mTeSR™3D media optimized for fed-batch culture of hPSC aggregates in suspension. A daily fed-batch culture system is employed, which eliminates the need for daily medium exchanges. By day 4 post-seeding, hPSC aggregates grow to a mean diameter of 400 µm, with a 5-fold increase in viable cell number. Confocal imaging of optically-cleared hPSC aggregates revealed uniform staining of hPSC markers and high viability. hPSC aggregates can be serially passaged by dissociating into small clumps using enzyme or enzyme-free methods, and re-seeding into new flasks. Use of STEMCELL Technologies STEMdiff™ Trilineage Differentiation Kit confirmed that cultures maintained in mTeSR™3D media differentiated into all 3 germ layers with high efficiency. Volumetric productivities were 0.7 and 6.9 (x105) cells/mL of medium used for standard 2D culture and fed-batch 3D cultures respectively. mTeSR™3D media used with an optimized fed-batch culture protocol enables efficient scale-up of hPSC production with greatly simplified workflow.
Robust and Consistent Conversion of Primed Human Pluripotent Stem Cells to Naïve-like Phenotypes
Friday June 24, 12pm-12:30pm | Level 2, Room 2009/2011Arwen Hunter, Scientist, STEMCELL Technologies
Traditional primed human pluripotent stem cell (hPSC) cultures correspond to late phase epiblast development and lack the core naïve state transcriptional circuitry observed in the mouse. Recent advances in the field have isolated and maintained naïve-like hPSCs with distinct developmental identities corresponding to earlier embryonic developmental phases. These advancements have allowed for the reversion of traditional primed hPSCs to these earlier states using specialized cell culture media and protocols. Naïve-like PSCs can subsequently be re-primed and maintained using traditional methods. This presentation will describe the application of our RSeT™ product line and protocols to robustly and consistently revert primed hPSCs to naïve-like states. RSeT™ products also allow for the continuous maintenance of the cells with phenotypic and karyotypic stability. We will examine the distinct characteristics of these cells including their core transcriptional profiles as well as the capacity to toggle between primed and naïve-like phenotypes. This tutorial will highlight STEMCELL Technologies’ integrated workflow for maintaining multiple pluripotent stem cell states and their subsequent downstream differentiation under defined culture conditions.
Conversion of Primed Pluripotent Stem Cells to a Naïve-Like State and Their Long-Term Maintenance Using RSet™ MediumArwen Hunter, Scientist – Poster board: #W2164 – Time: 19:30 – 20:30Brainphys™ Neuronal Medium: A Medium Optimized to Support the Synaptic Activity of Neurons Derived from Human Pluripotent Stem Cells and Primary CNS Tissues –
Vivian Lee, Senior Scientist – Poster board: #W2184 – Time: 19:30 – 20:30MesenCult™ Osteogenic Differentiation Medium – an Osteogenic Stimulatory Medium for the Efficient Differentiation of Human Mesenchymal Progenitor CellsArthur Sampaio, Senior Scientist – Poster board: #W1022 – Time: 19:30 – 20:30For more information about Stemcell Technologies’ Activities at ISSCR, please click here
Thermo Fisher Scientific – Booth 1725
Predict the unpredictable: using the in vivo niche to assess lineage potential and functionality of PSC-derived cellsThursday 23rd June, 8:00 – 8:30 AM, Floor 2, Room 2020Hideki Masaki, PhD, Institute of Medical Science, University of Tokyo
The lack of consistency in the differentiation and functional maturation of PSC-derived cells limits the application of these cells in translational research and regenerative medicine. One way to circumvent such issue is the usage of in vivo niche that provides the proper microenvironment to regulate cell fate. Our previous research demonstrated derivation of hematopoietic stem cells from PSCs via teratoma. We also reported the generation of whole organ from donor PSCs in allogeneic / xenogeneic organ-deficient animals. These proof-of-concept studies elucidated principles of a potential strategy for the design of stem-cell research and therapeutics. Recently, we discovered that by preventing apoptosis in the pre-implantation stage embryos we were able to form chimera from primed PSCs and even endodermal progenitors. In this presentation, we will discuss the potential impact of such in vivo system and its utility in accurately assessing lineage potential of stem cells and progenitor cells.
Part 1:Articlesized generation and characterization of patient-specific iPSC lines and gene editing of hESCs and iPSCsFriday 24th June, 11:30 – 12:30 PM, Floor 2, Room 2020Emily Titus, PhD, Manager, Technology, Centre for Commercialization of Regenerative Medicine (CCRM)
CCRM is a translational centre based in Toronto, Canada, focused on the development and commercialization of regenerative medicine and Cell Therapy technologies. An early achievement was the establishment of an iPSC production facility focused on generating high quality PSC lines for academic researchers and clinicians. Fully operational for three years, CCRM has delivered over 100 patient-derived iPSC lines currently used for disease modelling, and in drug screening initiatives, . Specializing in non-integrative reprogramming technologies, CCRM has developed SOPs to reprogram many common cell types in feeder-free conditions, including dermal fibroblasts, bone marrow stromal cells, cord and peripheral blood, and endothelial cells. iPSC lines are thoroughly characterized and tested using directed differentiation protocols to screen for pluripotent potential. Most recently, CCRM has established gene editing in hESCs and iPSCs using both TALENs and CRISPR/Cas9 to generate reporter cell lines and perform gene correction on patient-derived iPSCs. This presentation will focus on the technical challenges and associated solutions implemented at CCRM to establish these workflows.
Part 2:Neuronal differentiation of Parkinson’s patient-derived iPSCs into functional dopaminergic neuronsBirgitt Schuele, MD, Associate Professor, Program Director of Gene Discovery and Stem Cell Modeling, Parkinson’s Institute and Clinical Center
Differentiation into dopaminergic neurons that show midbrain specificity and exhibit specific cellular characteristics is critical for disease modeling, drug screening, and regenerative medicine. However, there are several technical hurdles such as yield of desired cell type, reproducibility of protocol, duration of differentiation, and cost. We compared Gibco Pluripotent Stem Cell (PSC) Dopaminergic Neuron Differentiation Kit (Prototype, Thermo Fisher Scientific, Cat. No. A30416SA) to our dopaminergic differentiation protocol (Mak et al. 2012) for length of protocol, efficiency of DA neuron generation, and electrophysiological properties. The Gibco PSC Dopaminergic Neuron Differentiation Kit showed a sharp increase in floorplate/mesencephalic markers such as FOXA2, CORIN, LMX1A, and EN1 at day 10 of neuronal induction. About 60% of total cells expressed the DA marker, tyrosine hydroxylase, after 35 days of differentiation and showed spontaneous activity on multielectrode arrays of ~3000 spikes and an average spike amplitude of 22µV (range of 17µV to 40µV). In summary, the Gibco PSC Dopaminergic Neuron Differentiation Kit provides reproducible conditions, shorter differentiation time, and a high-yield of functional dopaminergic neurons.
Development of a feeder-free PSC culture system enabling translational and clinical research
Wednesday, June 22, 2016: 19:30-20:30 Poster #: W3002
A robust platform for generation and high throughput functional analysis of human induced pluripotent stem cell-derived cardiomyocytes
Thursday, June 23, 2016: 18:00-19:00 Poster # T3029
Recombinant human laminin-521 supports PSC survival in Essential 8 Media during critical transitions
Thursday, June 23, 2016: 18:00-19:00 Poster #: T3021
Optimization of human CD34+ cells for reprogramming using tailored media and alternate timelines
Thursday, June 23, 2016: 19:00-20:00 Poster #: T2102
Generation of large numbers of floor plate derived, midbrain-specified DA neurons from human PSC for scaled applications
Friday, June 24, 2016: 18:00-19:00 Poster #: F2035
Advanced delivery technologies for improved CRISPR-based genomic editing in stem cells and induced pluripotent stem cells for disease model generation
Friday, June 24, 2016: 19:00-20:00 Poster #: F3002
Learning lab and lounge:
Stop by the Thermo Fisher Scientific training lab and lounge for a chance to relax and refresh your Stem Cell Research skills. Join us as we discuss some of the most common questions researchers ask and learn how to optimize your cell culture system. Each stop in the training lab will take 10 minutes – attend all 6 for entry into a raffle drawing for a Gibco Education PSC Prize Package.*
Meet Up Hub:
Let’s Talk Clinical Translation
Thursday, 23 June, 3:15-4:00PM – Exhibition Hall – Meet-up Hub 1
Connect with your colleagues around the world to discuss your Cell Therapy clinical translational endeavors and share best practices and challenges. Let’s talk about how to design a pre-clinical roadmap, common considerations and useful tips to help expedite your path to the clinic. This hub is organized by Thermo Fisher Scientific.
For more information about Thermo Fisher Scientific’s Activities at ISSCR, please click here