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CELLutions SUMMIT - Day 1

Cell Sources

Unlike organs, cells are a potentially renewable resource for body repair. The initial excitement generated by the identification of novel cell sources must give way to focused efforts on methods to manipulate their differentiation and self-renewal capabilities. The best thing is to have a variety of cell sources to provide the best cell for potential patient therapy. The focus is on the development of technologies to isolate, culture, manipulate, and differentiate cells.

SUNDAY, AUGUST 10

5:00-6:00 pm  Early Registration

MONDAY, AUGUST 11

7:30-8:30 am  Registration and Morning Coffee

iPS CELLS

8:30 Chairperson’s Opening Remarks
David Kaplan, Ph.D., Professor, Biomedical Engineering, Tufts University

8:40  Generation and Potential Applications of Human Induced Pluripotent Stem Cells from Dermal Fibroblasts
William E. Lowry, Ph.D., Assistant Professor, Department of Cellular and Development Biology, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, UCLA
The newest method to potentially generate patient-specific stem cells was introduced in 2006 with a landmark study showing that mouse fibroblasts could be reprogrammed simply by introduction of defined factors.  Recent efforts to reprogram cells by induction of defined factors is an extrapolation of the simple, yet elegant idea that cells might have the ability to change their fate if their gene expression profile is manipulated to reflect that of another cell type.  In the case of murine iPS, several groups found that a group of genes that is highly expressed in mESCs could be transduced into fibroblasts, and that these genes would then reprogram the fibroblast to an embryonic state.  Four groups, including ours, have now shown that this general method can be used to reprogram human fibroblasts,  The ideas that cells with all the properties of HESCs could be generated from easily accessible fibroblasts has changed the way we think about stem cells and their potential application in clinical setting.  We introduced OCT4, SOX2, KLF4, and C-MYC into neonatal human fibroblasts and found that, similar to what was shown in the murine system, human fibroblasts could be converted to an embryonic state.  Despite having undergone a massive reorganization of their epigenetic and transcriptional landscape, none of the lines generated in our lab have displayed any gross chromosomal abnormalities.  In addition, these cells are thriving after 30 passages to date and have retained their ability to differentiate down lineages representative of the three embryonic germ layers in vivo and in vitro.  These human iPS cells seem to have all the potential of human embryonic stem cells and can be derived from the progeny of cells in a simple skin biopsy obviating the need for human embryos and opening the door to the possibility of patient specific stem cells.

9:10 Production of iPS Cells from Somatic Cells of Patients with Genetic Disease
In-Hyun Park, Ph.D, Research Fellow, Department of Hematology and Oncology, Children’s  Hospital Boston
We obtained fibroblast cultures from patients with a variety of genetic diseases, and subjected them to infection with retroviruses expressing the four transcription factors. After growth and selection for 3-4 weeks, we isolated and characterized iPS colonies. Morphologically, disease-specific iPS cells appear indistinguishable from hES cells and iPS cells from normal somatic cells.  The iPS cells and hES cells share a similar expression pattern of pluripotent genes and surface markers. When differentiated as embryoid bodies in vitro, they express genes for all three germ layers. When injected subcutaneously into immune-compromised mice, they form well-defined teratomas containing tissues from all three germ layers, thereby demonstrating their pluripotency. Disease-specific iPS cells represent a substrate for experimental analysis of disease pathophysiology, a reagent for cell-based drug screening, and a target for gene repair. Disease-specific iPS cells represent an important new tool for stem cell research.

9:40  Pluripotent Stem Cell Culture Media Considerations
Todd McDevitt, Ph.D., Assistant Professor, Biomedical Engineering, Georgia Institute of Technology 
Pluripotent stem cells are exquisitely sensitive to environmental cues and chiefly among them, soluble media components are critically important for proper maintenance and propagation of the cells.  A number of individual soluble factors and combinations of such molecules used successfully to propagate pluripotent stem cells have begun to be developed into commercially available products for pluripotent stem cell culture.  Issues associated with the sources of soluble materials, quality control metrics, synergy with matrix components and applicability to different pluripotent cell types will be discussed.

10:10  Networking Coffee Break

CELL SOURCES

10:30  Technology Spotlight (Sponsorship Available)

10:45  A Pure Ectodermal Cell Population Derived from Human Embryonic Stem Cells for Skin and Cornea Cell Therapy
Daniel Aberdam, Ph.D., Director, Stem Cells, Development and Cancer Department, INSERM and Director, Rappaport Institute, INSERTECH 
Among the different cell types that can be generated from ES cells, ectodermal cells are of particular interest since they represent the precursors of many epithelial tissues and organs. Here we report the reproducible isolation of an ectodermal cell population from human ES cells (IT1). IT1 shows a stable morphological appearance, has a somatic cell cycle kinetics and does not produce teratoma in nude mice. Remarkably, like adult somatic cells, IT1 cells undergo senescence after about 60 doubling population and their karyotype remains normal. Exogenous expression of p63 or Pax-6 efficiently induces their differentiation into K14+-keratinocytes or K12+-corneal cells, respectively. In addition, this multipotency of IT1 is confirmed on defined extra cellular matrices. IT1 cell line could be thus a particularly useful tool to design in vitro models that would recapitulate the embryonic early events of epithelial lineage specification but also as a source of different cells for clinical trials of epithelial stem cell loss. This reproducible protocol could easily be transposed to any pluripotent cells, including the newly discovered iPS.

11:15  Building a Better Hepatocyte Model: Human Stem Cell-Derived Hepatocytes 
Marsha Roach, Ph.D., Vice President, Research, CellDesign, Inc. 
To address the need for physiologically-relevant human hepatocytes, multiple surrogate hepatic model systems, such as liver microsomes, cryopreserved hepatocytes and HepG2 cells have become the current standards in the pharmaceutical industry; however, all of these models have limitations.   In order to develop a better system that has more complete hepatic function, methods are being developed to direct the in vitro differentiation (IVD) of stem cells into hepatocytes that may be more functional, more physiologically relevant and more predictive of clinical outcome than current models, therefore reducing attrition of drugs that enter the clinic.   Here we describe a staged process for the efficient and reproducible differentiation of stem cells into mature, functional hepatocytes and provide preliminary data to demonstrate that stem cell-derived hepatocytes can be used  as a in vitro system for toxicity screening in drug discovery.

11:45  Patient-Specific HLA Homozygous and Heterozygous Stem Cell Lines Derived from Human Parthenogenetic Blastocysts
Nikolay Turovets, Ph.D., Director, Research and Therapeutic Development, International Stem Cell Corporation
Parthenogenetic activation of human oocytes may be one way to produce histocompatible cells for cell-based therapy. Individual HLA homozygous parthenogenetic human stem cell (hpSC-Hhom) lines have the potential for cell-based therapy in a significant number of individuals, provided the HLA haplotype is prevalent. We report the successful derivation of four stable hpSC-Hhom lines from both HLA homozygous and HLA heterozygous donors as well as six patient specific pluripotent human embryonic stem cell (phESC) lines from blastocysts of parthenogenetic origin. All lines demonstrate typical human embryonic stem cell morphology, express appropriate stem cell markers and possess high levels of alkaline phosphatase and telomerase activity. Additionally, injection of these cell lines into immunodeficient animals leads to teratoma formation. HLA genotyping of all four hpSC-Hhom lines demonstrates that they are HLA homozygous. Single-nucleotide polymorphism (SNP) data analysis suggests that hpSC-Hhom lines derived from HLA heterozygous oocyte donors are homozygous throughout the genome. DNA profiling of all six phESC lines demonstrates that they are MHC matched with the oocyte donors.

12:15 pm  Close of Session

12:30  Luncheon Technology Workshop

Sponsored by
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Engineering Zinc Finger Proteins (ZFPs) for Stem Cell Research and Therapy
Trevor Collingwood, Ph.D., Team Leader, Cell Engineering, Sangamo Biosciences, Inc. 
Zinc finger proteins (ZFPs) are the dominant class of naturally occurring DNA-binding motifs found in nature. Attaching different transcription regulatory domains to engineered site-specific ZFPs enables us to achieve the distinct effects of targeted endogenous gene activation or repression. Using this powerful platform, ZFPs have been designed to promote the mobilization of endogenous stem cells, thus offering new strategies for cell regeneration and repair. Moreover, by linking a nuclease domain to ZFPs, the resultant zinc finger nucleases (ZFNs) enable highly efficient precision genome editing which results in targeted knock out, correction or insertion of investigator-specified genes in human stem cells. Thus, ZFPs represent a powerful tool for stem cell research and therapy.

1:45 Session Break

INTERACTION OF CELLS AND MATERIALS

2:00  Chairperson’s Remarks
Dolores Baksh, Ph.D., Senior Scientist, Research & Development, Organogenesis Inc.

2:05  Stem Cells and Materials for Vascular Differentiation and Regeneration
Sharon Gerecht, Ph.D., Assistant Professor, Chemical and Biomolecular Engineering, Johns Hopkins University
Vasculogenesis is the process in which blood vessel formation from vascular endothelial cells occurs by the differentiation of precursors, which in turn give rise to a primitive vascular plexus. The goal of our studies is to utilize engineered biomaterials to understand the mechanisms underlying microenvironment regulatory cascades during vascular differentiation and regeneration.

2:35  Modeling Human Skin in Structure and Function: The Importance of Cross-Talk between Fibroblast and Keratinocytes and Extracellular Matrix
Dolores Baksh, Ph.D., Senior Scientist, Research & Development, Organogenesis Inc.
The successful maturation and function of tissue-engineered skin is dependent on the orchestrated cross-talk between fibroblasts and keratinocytes, where fibroblasts provide the necessary growth factor and extracellular matrix signals that support keratinocyte growth, terminal differentiation and survival. Organogenesis Inc. has developed a commercially available tissue engineered skin equivalent, Apligraf (R), which approximates human skin in structure and barrier function.  Apligraf(R) incorporates a layer of fibroblasts seeded in a biomaterial under a sheet of keratinocytes. The resultant living skin equivalent displays phenotypic characteristics of healing skin, incorporating matrix proteins and cytokines found in human skin.  The tissue engineered skin has already demonstrated clinical success in the treatment of diabetic foot ulcers and venous leg ulcers. On-going research efforts are focused on elucidating the bioactive components of the living skin construct in supporting wound healing.

3:05  An FGF-2-Modified Nanofibrillar Prosthetic for Spinal Cord Repair
Sally Meiners, Ph.D., Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School 
We have developed a spinal cord prosthetic (SCP) that incorporates narrow strips of randomly oriented nanofibers that are longitudinally bundled to provide appropriate geometric cues for axonal regrowth.  Our data suggest that the SCP can correctly guide regenerating axons across the injury gap created by an over-hemisection of the adult rat thoracic spinal cord.  In addition, devices that incorporated nanofibers modified with fibroblast growth factor-2 (FGF-2) encouraged substantially more axonal regeneration and better functional recovery (assessed using the Basso, Beattie, Bresnahan (BBB) locomotor rating scale) than did devices that incorporated unmodified nanofibers.  Furthermore, an FGF-2-modified SCP also encouraged revascularization but did not promote glial scarring or an apparent foreign body or inflammatory response.  As such, the FGF-2-modified SCP provides a multi-faceted approach to spinal cord repair.

3:35  Networking Refreshment Break

FIVE @ FIVE
Building Cell-Based Breakthroughs into a Viable Business

Session Chairperson: Annette Reynolds, Executive Director, 128 Innovation Capital Group

4:00  VC Panel of Experts
Anthony G. Coia, M.D., BioVentures Investors
Jim Sherblom, MBA, Managing General Partner, Seaflower Ventures
Leonide Saad, Ph.D., Proteus Venture Partners

The Five @ Five Emerging Technology Showcase features a unique format that consists of:

  • 5 minute “elevator” presentations
  • 5 powerpoint slides
  • 5 emerging companies

4:40 Primorigen Biosciences, LLC
Spots-On-Dots: A Novel Frameless Microarray Platform for Multiplexed Biomarker Profiling
Nicholas Caruccio, Ph.D., Director of Marketing

4:50 Capricor Inc.
Cardiac Stem Cell Therapies for Heart Disease
Rachel Ruckdeschel Smith, Ph.D., Senior Scientist & Project Lead, Regnerative Medicine

5:00 Tissue Regeneration Therapeutics Inc.
HUCPVC Technologies
John E. Davies, Ph.D., President

5:10 Cell & Tissue Systems, Inc.
Preservation Strategies for Cell and Tissue Therapeutic Products
Kelvin Brockbank, Ph.D., President and CSO

5:20 Bionas
A Novel Cell-Based Analyzing System for Continuous Determination of Cell Physiology
Carsten Haber, Ph.D.

5:30  Grand Opening of Exhibit Hall and Networking Reception

7:00  Close of Day 

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