Thursday, June 19, 8:00 a.m. - 8:30 a.m., West Ballroom A
An Integrated Workflow for the Isolation, Expansion, and Characterization of Hematopoietic Progenitors and their Conversion to Induced Pluripotent Stem Cells in Defined Culture Conditions
Wing Chang, PhD, Scientist and Steve Szilvassy, PhD, Director, Hematopoietic Products
Hematopoietic stem and progenitor cells are undoubtedly the most well studied tissue-specific precursor cells and now are commonly used for generating iPSCs. This presentation will describe the isolation, expansion, and characterization of human blood-derived CD34+ progenitors and erythroid cells, and methods for subsequent reprogramming of these cells to iPSCs using defined culture conditions. The tutorial will highlight STEMCELL Technologies integrated workflow for manipulating hematopoietic development in vitro and the generation of iPSCs from these precursors.
Thursday, June 19, 8:00 a.m. - 8:30 a.m., West Ballroom B
Differentiating Stem Cell Populations with Success
Joy Aho, PhD
The utility of stem cells resides in their ability to differentiate into a multitude of cell types, providing powerful tools for regenerative medicine and drug toxicity testing. Differentiation protocols can be fraught with unknown variables and are time- and reagent-consuming. We present tools and protocols to reduce experimental variability and drive efficient stem cell differentiation. We will discuss methods to simultaneously measure protein levels of established markers and investigate functional characteristics of derivative cells.
Thursday, June 19, 8:00 a.m. - 8:30 a.m., West Meeting Room 211-214
Large particle flow cytometry provides high throughput analysis and automation for cell clusters (EBs, spheroids) and encapsulated 3D cell cultures.
Rock Pulak, Ph.D.
Embryonic stem cells (embryoid bodies), neural stem cells (neurospheres), certain solid tumors (tumorspheres) and other cell types all tend to form cell clusters. These clusters are a more natural arrangement, where cell-cell interactions and cell-substrate contacts can be established and maintained. Union Biometrica discusses the use of their Large Particle Flow Cytometers for automating the multiparametric analysis and gentle intact dispensing of these types of cell clusters to wells of multi-well plates.
Thursday, June 19, 8:00 a.m. - 8:30 a.m., West Meeting Room 301-305
Nikon Corporation Using Stem cells to study Neurodegenerative Disease
Prof. Lee Rubin, Harvard University
The general idea of using stem cells to discover new treatments for serious neurodegenerative diseases, especially, “in vitro clinical trial” using the live cell imaging system is described. Nikon has been provided the live cell imaging system (BioStation CT) and it has been well accepted in the stem cell research field. This advanced system has the capability of the long term observation and measuring cell behavior with image analysis technology.
Thursday, June 19, 11:30 a.m. - 12:30 p.m., West Ballroom A
STEMCELL Technologies Induction of a Native State in Human Pluripotent Stem Cells
Huck-Hui Ng, PhD, Executive Director, Genome Institute of Singapore
In contrast to traditional hPSC cultures, native or naïve hPSCs are more primitive, resembling cells of the pre-implantation blastocyst. Beyond their importance for studying early development, naïve cells may accelerate pluripotent cell therapies, owing to their single-cell passaging compatibility, robust growth and amenability to genetic manipulation. Dr. Ng will discuss his ground-breaking research on induction and maintenance of a novel state in established human embryonic stem cell lines using mTeSR™1 medium and small molecules.
During the second half-hour, from 12:00-12:30pm, Erik Hadley, PhD, Senior Scientist
will present Scalable Enzyme-Free Protocols for the Isolation and Maintenance of Human Induced Pluripotent Stem Cells (hiPSCs) Without Mechanical Colony Scraping
. This tutorial focuses on fibroblast reprogramming and hiPSC maintenance protocols which use ReLeSR™, an enzyme-free passaging reagent that eliminates manual removal of differentiated cells, colony scraping and complicated techniques to obtain uniform cell aggregates. When used with mTeSR™1 or TeSR™-E8™, ReLeSR™ enables the use of closed vessels, thus facilitating scale-up and automation. Applicability of ReLeSR™ in a complete workflow from fibroblast reprogramming to the establishment of a new hiPSC line will be discussed.
Thursday, June 19, 11:30 a.m. - 12:30 p.m., West Ballroom B
LONZA Bridging Research to Therapy: Novel iPSC Tools and Technologies
Behnam Ahmadian Baghbaderani, Ph.D., Lonza Cell Therapy Development Services
Dr. Yamanaka’s Nobel Prize winning discovery has spurred scientific advancements in basic research, disease modeling, drug development, and regenerative medicine. We have assembled three speakers that describe how Lonza is working to help stem cell researchers overcome their challenges.
From the Nobel Prize to the Clinic: One Step Closer to iPSC-based TherapiesDr. Baghbaderani
Behnam Ahmadian Baghbaderani, Ph.D., Lonza Cell Therapy Development Services
Lonza L7™ Reprogramming and Cell Culture System - A Case Study
Yu-Chieh Wang, Ph.D., Center for Regenerative Medicine, The Scripps Research Institute
Highly Efficient and Specific Human Genome Editing in iPSCs Using CRISPR-Cas9 System
Linzhao Cheng, Ph.D., Professor of Medicine and Oncology, Stem Cell Program, Johns Hopkins School of Medicine
will introduce Lonza’s L7™ Reprogramming and Culture System outlining the benefits for the generation and maintenance of iPSCs for research and clinical applications. Dr. Wang
will review the potential challenges in cell reprogramming and how the L7™ System helps researchers overcome these hurdles. Lastly, Professor Cheng
will present the latest results on human genome editing in iPSCs using CRISPR-Cas9 system and address issues related to the level of off-target rates in human iPSCs. Collectively, these timely presentations will showcase the latest innovative technologies and tools to empower scientists conducting human stem cell research.
Thursday, June 19, 11:30 a.m. - 12:30 p.m., West Meeting Room 211-214 Dissecting developmental trajectories using single-cell transcriptional profiling
Craig E. Nelson, Ph.D., Associate Professor Molecular & Cell Biology University of Connecticut
Advances in microfluidic technology have moved single-cell biology into the mainstream. Here we will present the application of single cell transcriptional profiling to dissecting complex developmental events controlled by wild-type, mutant, or ectopic genomes.
Common measures of gene expression lose information because they only operate at the population level: the results can only represent the strict average of the population. As a result many subtleties are blurred; decision points in cell lineages and the beginnings of differentiation, and the co-regulation of genes in individual cells are often lost in population-level measurements. Over the last several years we have developed a suite of methods and computational tools to to support the robust qualitative and quantitative measurement and analysis of transcription in single cells. We will be presenting these tools in the context of understanding critical events in embryogenesis, differentiation, and the reprogramming of somatic cells to pluripotency.
Thursday, June 19, 11:30 a.m. - 12:30 p.m., West Ballroom CD Cell Surface Marker Screening & Analysis of Stem Cell Populations
Robert Balderas and Christian Carson
The first half-hour will be dedicated to how cell surface marker analysis of stem cells allows for phenotypic characterization as wll as cell purification. This tutorial will include the disscussion of the following topics:
- Designing and performing cell surface marker screens to identify unique surface signatures for the isolation of stem cells and their derivatives
- Effective sorting of pluripotent and neural lineages
- Flow Cytometry panel design and instrument optimization for stem cell populations
The second half-hour, from 12:00-12:30 p.m., will focus on the Use of Intracellular Flow Cytometry to Monitor Lineage Differentiation and Potency in Stem Cells
. Presenters Robert Balderas
and Nil Emre
will demonstrate how intracellular flow cytometry enables the quantitative measurement of stem cell populations and their derivatives. Applications discussed will include:
- Effective cell preparation and procedural considerations when performing intracellular flow cytometry
- Analysis of neural, cardiac, and endodertm differentiation of hESCs using intracellular flow cytomety
- Assays to measure immunomodulation by MSCs
Thursday, June 19, 11:30 a.m. - 12:30 p.m., West Meeting Room 301-305
Reprogramming somatic cells using the Sendai virus technology
Laurence Daheron, Harvard University
In recent years, non-integrating reprogramming technologies have been developed to efficiently generate safer iPS lines. One of them uses the Sendai virus as a vector to introduce the four Yamanaka factors into somatic cells. The Sendai virus is a single stranded RNA virus that can effectively transduce a variety of cells. At the HSCI iPS core, we’ve widely used this method to derive induced pluripotent stem cells. We will discuss the advantages of this method.
In the second half-hour from 12:00-12:30pm, Dr. Dhruv Sareen
, Director, iPSC Core at Cedars-Sinai Medical Center will present, From bland to grand: iPSC technology and its applications
. A significant bottleneck in drug development is reproducible and unlimited supply of physiologically-relevant human cells that bear patient-specific genetics. Human iPSC technology promises to circumvent this and support development of more predictive high-content cellular assays. I will highlight reprogramming and differentiation technologies used in the CSMC iPSC core with emphasis on:
- Platforms for high-content screening/imaging (HCS/HCI)
- Developing predictive cellular assays with iPSCs
- iPSC disease modeling with HCI
- Challenges for translating iPSC-derivatives from lab-scale to high-content
Friday, June 20, 8:00 a.m. - 8:30 a.m., West Ballroom A Introducing a Novel, Animal Component-Free (ACF) Culture System for Efficient Isolation, Expansion and Cryopreservation of Human MSCs Derived from Bone Marrow and Adipose Tissue
Ravenska Wagey, PhD, Senior Scientist
This tutorial will describe MesenCult™-ACF, a novel animal component-free culture system optimized for efficient isolation, expansion and cryopreservation of human MSCs (hMSCs) derived from primary tissues. Clonogenic characteristics, phenotype and long-term expansion of hMSCs under strict ACF culture conditions will be discussed. The application of three robust and versatile differentiation kits to characterize these hMSCs will also be introduced, providing a complete and integrated workflow for efficient isolation, expansion, differentiation and cryopreservation of hMSCs.
Friday, June 20, 8:00 a.m. - 8:30 a.m., West Ballroom B
Next-generation technologies for stem cell research
Sebastian Knoebel, PhD, Senior Project Manager R&D Stem Cells
The development of clinically compliant technologies is key in transitioning from basic stem cell research to clinical applications. We will introduce workflows for the integration-free reprogramming and modulation of stem cells using modified mRNAs. Furthermore, we will present a novel xeno-free culture system that guarantees robust, feeder-free cultivation of human pluripotent stem cells. Lastly, the microchip-based MACSQuant® Tyto enables gentle, multiparameter flow sorting in a sterile, closed system complementing our MACS® Separation Technology.
Friday, June 20, 8:00 a.m. - 8:30 a.m., West Ballroom C/D The Laminin Protein Family – The key to primary cell culture difficulties
Kristian Tryggvason, PhD, MBA
Primary cell culture has four major problems. First, expansion of pluripotent cells has been considered difficult. Second, keeping adult differentiated cells in their differentiated state in vitro for prolonged periods was previously impossible. Third, differentiating primary cells on biorelevant matrices is a large challenge. And fourth, it has been considered impossible to culture adult cells or tissue samples. With the help of the Laminin protein products, we have solved all these problems.
Friday, June 20, 8:00 a.m. - 8:30 a.m., West Meeting Room 211-214 Optimizing the culture of PPSC using novel Xeno and Insulin Free Low Protein Media
Rick.I. Cohen, Ph.D., Rutgers
This tutorial will introduce Animal Free PeproGrow-hESC, a new Pluripotent Stem Cell Media with companion products and the proper techniques to optimize the growth iPSC or hESC. Our presentation will include, but not limited to; the use of this novel media during iPSC generation; Cryopreservation and Thawing, Growth VitroGrow-hESC (recombinant full length human Vitronectin), and ability to passage cells at wildly high dilution factors.
Friday, June 20, 8:00 a.m. - 8:30 a.m., West Meeting Room 301-305 HumaXpress Human Cell Expression System
Dr. Mark Azam
HumanZyme has developed a unique HEK293 expression system to express and purify human cytokines using serum- and protein-free, chemically defined media. This streamlined and scalable system produces human proteins that have native folding, disulfide bridge formation, proteolytic processing, and glycosylation. The advantages of this system to produce human proteins with higher stability, consistency, and biologic activity will be discussed.
Friday, June 20, 11:30 a.m. - 12:30 p.m., West Ballroom A
Biological Industries Israel Beit Haemek Ltd.
A novel xeno-free culture system for isolotation and expansion of hMSC from various sources toward cell therapy applications
hMSC serves as a new, promising tool for regenerative medicine and cell therapy with advantages over other stem cells types, mainly due to their multipotent characteristic, broad variety of tissue sources and immuno-privilege. The presentation addresses the ability of MSC Nutristem XF culture medium as well as auxiliary solutions for attachment, dissociation, and cryopreservation to isolate hMSC from various tissue sources and to support long-term expansion of multipotent hMSC suitable for cell therapy applications.
Friday, June 20, 11:30 a.m. - 12:30 p.m., West Ballroom B
Advanced Technologies for in vitro culture of cell types relevant for clinical research
Zara Melkoumian, PhD
Today's complex cell culture techniques, such as stem cell culture and tissue engineering, require in vitro environments with expanded capabilities. These include xeno-free and human-origin component-free culture conditions and scalable closed vessel design. Corning brings together a comprehensive, innovative line of vessels, media and surfaces designed and characterized to meet the needs of these advanced applications, enabling researchers with the right environment for every cell. This tutorial will focus on technologies available for expansion and functionality of clinically relevant cell types including, mesenchymal stem cells (MSCs), human keratinocytes (HKN), endothelial colony forming cells (ECFCs), neural progenitor cells (NPCs) and other attachment dependent cell types.
In the second half-hour from 12:00-12:30pm, Deepa Saxena, PhD
will present Surface Applications for Feeder-free Culture of Human Pluripotent Stem Cells (HPSC)
. Human pluripotent stem cells have an infinite capacity of self-renewal and differentiation into all derivatives of the three germ layers. A great deal of hope is associated with the potential application of these cells in functional genomics, cell therapy, and regenerative medicine. Conventionally, hpsc have been cultured on mitotically inactivated murine embryonic fibroblast (MEF) cell feeder layers. Robust, easy to use, reproducible and scalable culture systems are desired for undifferentiated expansion of these cells. Corning® offers several matrices that support feeder-free hPSC expansion. Tutorial will provide an overview of natural and recombinant/synthetic matrices for hPSC culture.
Friday, June 20, 11:30 a.m. - 12:30 p.m., West Ballroom C/D
EMD Millipore Corporation
Virus-free, efficient reprogramming using a single transfection of a synthetic, polycistronic self-replicating RNA
Vi Chu, Ph.D. Manager II, R&D, Stem Cells/Cell Biology, EMD Millipore
Drawbacks of current iPSC generation methods, involving virus or multiple transfections, include experimental variability, questions of biosafety, and known and unknown effects of introducing viruses into experimental systems. We describe a simple, non-integrating reprogramming system in which a single transfection of a synthetic, polycistronic (OKS-Glis), self-replicating RNA species was sufficient to generate high numbers of human iPSCs. The efficiency of this new RNA-Replicon Reprogramming Technology ranged from 0.3% to 1.1%, depending on fibroblast proliferation rate.
Friday, June 20, 11:30 a.m. - 12:30 p.m., West Meeting Room 211-214
Irvine Scientific Ex Vivo Expansion, Differentiation and Cryopreservation of Mesenchymal Stromal/ Stem Cells
Ning Liu, PhD
Mesenchymal stromal/ stem cells (MSCs) are capable of self-renewal, differentiation into various cell lineages, and immunomodulation, which make them a valuable source of cells for clinical applications. This workshop will illustrate the development of a portfolio of serum-free, ready-to-use and customizable media for ex vivo expansion, differentiation and cryopreservation of human MSCs. It will demonstrate the superior performance of our cGMP grade medium and how it can facilitate consistent and high quality clinical research of MSCs.
Friday, June 20, 11:30 a.m. - 12:30 p.m., West Meeting Room 301-305
RNA-mediated generation of integration-free iPS cell lines from cells isolated from human blood
Brad Hamilton, Director of Research and Development
Stemgent-Asterand’s non-integrating mRNA reprogramming platform utilizes microRNA to facilitate fast and efficient establishment of iPS cell lines from diseased human fibroblasts that often times can be refractory to other reprogramming methods. Here we present data highlighting the application of novel RNA reprogramming technologies that, in combination with microRNA, enable the generation of stable, fully reprogrammed iPS cell lines from late-outgrowth endothelial progenitor cells (L-EPCs) isolated from human peripheral blood.
Saturday, June 21, 8:00 a.m. - 8:30 a.m., West Ballroom A
LifeMap Sciences Inc.
LifeMap Discovery® - the Roadmap for Stem Cell Research
Idit Livnat, PhD
LifeMap Discovery (http://discovery.lifemapsc.com) is a powerful stem cell research platform that provides comprehensive mapping of embryonic development along with substantial information about stem and progenitor cells, their differentiation protocols and cell therapy applications. We will showcase our platform and demonstrate its value for stem cell research and the development of therapeutic products. GeneAnalytics, our novel tool for classification and identification of derived stem cells, will also be presented.
Saturday, June 21, 8:00 a.m. - 8:30 a.m., West Ballroom B
EMD Millipore Corporation
Expansion and Harvest of Adult Stem Cells Supports Large Scale Manufacturing
As more stem cell therapeutics progress through clinical testing, current in vitro culture methods are cumbersome to scale. In this case study, we verified that cells expanded in the single use stirred tank bioreactor and subsequently harvested were identical in phenotypic profile in comparison to flat culture and maintained the desired cell characteristics of hMSCs, thereby confirming the consistency, quality and reproducibility of large scale in vitro systems for stem cell expansion.