Molecular Embryology & Stem Cell Biology
Embryonic, Pluripotent & Heamotopoietic Stem Cells
Stem Cells & Policymaking & Political Perspectives
Epigenetics of Stem Cells and Cell Imaging
Control of Cell Differentiation, Self-renewal
Reprogramming and Stemness redux
Adult Stem Cells & Stem Cell Replacement Therapy
Regenerative Medicine & Intellectual Property Issues
Biopolymers and Tissue Engineering
Stem Cell Biotechnology: Cell Culture, and Expansion Methodologies
Advances in Stem Cell Therapeutics
Panel Discussion with experts from VC firms, IP Law Firms and Media
SPECIAL LECTURE on Bioethics

Highlights of the meeting:
Inaugural Address by Nobel Laureate Dr. P. Berg of Stanford
Keynote by Embryonic Stem Cell Pioneer Dr. G. Martin of UCSF
Stem Cell Legislation by California Democrat State Senator D. Oritz
Bioethics Lecture by US President’s Ethics Committee Member Dr. W. Hurlbut
Pluripotent Stem Cell Therapies for Ischemic Injury and Diabetes
Lineage Selection from Differentiating Transgenic ES Cell Culture
Human Embryonic Stem Cell-based Therapy for Parkinson’s disease
Adult Neural Progenitor Cell Replacement Therapy for Huntington’s disease
Angiogenic Progenitor Cell Transplantation for Cardio-myopathy
Advances in Cardiac Stem Cell Therapy and Transplantation
Role of MicroRNAs during Cardiac Progenitor Differentiation and Cardiogenesis
Adult-Derived Hepatic Stem & Neural Stem Cells for Clinical Applications
Use of Cord Blood Stem Cells in Tissue Engineering of the Eye
3-Dimensional Biodegradable Scaffolds for Embryonic Stem Cell Growth
Controlling Stem Cell Fate via Biomimetic Polymers
Stem Cell Scale-up in Bioreactors and how to Manufacture Therapeutic Products
Somatic Cell Cloning & Nuclear Reprogramming
How will Intellectual Property Factors Affect the Commercialization of Stem Cell Therapeutics?
Stem Cells, Embryos and Ethics: Is There A Way Forward?

AN EXCELLENT MEETING TO MEET WORLD CLASS SCIENTISTS, POLICY MAKERS & BIOETHICISTS

Paul Berg, PhD. Nobel Laureate 1980 (Chemistry) Cahill Professor of Biochemistry, Emeritus Stanford University School of Medicine Stanford, California, 94305 USA Title: "Brilliant Science-Dark Politics"

Keynote Speaker (Stem Cell Innovator Award)

Gail Martin, PhD. Professor & Head Program in Developmental Biology Department of Anatomy at School of Medicine University of California at San Francisco, San Francisco, CA 94158

Key speakers includes (see below for abstracts):

Yerem Yeghiazarians, MD.
Assistant Professor of Medicine & Director of Cardiac Stem Cell Translational Development Program
University of California at San Francisco Medical Center
Division of Cardiology, San Francisco, CA 94143-0124
Title: Advances in Cardic Stem Cell Therapy

Bronwen Connor, PhD.
Senior Lecturer & Head of the Neural Repair and Neurogenesis Lab
University of Auckland, New Zealand
Title: Adult Neural Progenitor Cells and Cell Replacement Therapy for Huntington's Disease

G. Rasul Chaudhry, PhD.
Professor , Department of Biological Sciences, Oakland University, Rochester, MI
Title: Tissue Engineering Potential of Embryonic Stem Cells Using Three-Dimensional
Biodegradable Scaffolds

Kevin Healy, Ph.D.
Professor, Departments of Bioengineering and Materials Science & Engineering
University of California at Berkeley, Berkeley, CA USA
Title: Controlling Stem Cell Fate via Biomimetic Polymers

Todd N. Spalding
Associate of the Life Sciences Industry, Foley & Lardner LLP., Washington D.C.
Title: How Will Intellectual Property Factors Affect the Commercialization of Stem Cell Therapeutics?

Daniel Kraft, MD.
Senior Fellow, Pediatric Hematology/Oncology, Stanford University School of Medicine, Stanford, CA 94305 USA
Title: Hematopoietic Stem Cells: From Bench to Bedside

Evan Snyder, MD., PhD.
Professor & Program Director, Stem Cell Research Center, The Burnham Institute, La Jolla, CA
Title: Assessing the Genetic and Epigenetic Stability of Stem Cells

Ana Sofia Correia
PhD student (from Professor Patrik Brundin’s Lab)
Faculty of Medicine, Lund University
Department of Experimental Medical Science
Wallenberg Neuroscience Center, Lund, Sweden
Title: Developing Human Embryonic Stem Cell-based Therapy for Parkinson’s disease

Xiangzhong (Jerry) Yang, PhD.
Professor & Director, Center for Regenerative Biology, University of Connecticut Storrs, CT
Title: Somatic Cell Cloning and Nuclear Reprogramming via Cloning from de differentiation to re-differentiation – State of the Technology and Future Applications

Michael Mann, MD.
Asst. Professor of Surgery, University of California at San Francisco, San Francisco, CA
Title: Cardiac Stem Cell Transplantation: The Rest of the Story

David T. Harris, PhD.
Professor
Department of Microbiology and Immunology
Director, Stem Cell Bank, University of Arizona, Tucson, AZ
Title: The Potential of Cord Blood Stem Cells for Use in Tissue Engineering of the Eye

Deepak Srivastava, MD
Professor & Director Gladstone Institute of Cardiovascular Disease
University of California San Francisco, San Francisco, CA 94158
Title: Role of microRNAs during cardiac progenitor differentiation and cardiogenesis

Dr. Nicole I. zur Nieden, PhD.
Post doctoral Fellow in the lab of Dr. Derrick E. Rancourt
Institute of Maternal & Child Health, University of Calgary, Calgary, AB, CANADA
Title: Billions of embryonic stem cells for cytotherapy: quality controlled expansion in bioreactors

Robert J Deans, PhD.
Vice President of Regenerative Medicine, Athersys, Inc., Cleveland, OH
Title: Development of Pluripotent Stem Cell Therapies for Ischemic Injury and Disease

Anish S. Majumdar, PhD.
Senior Director of Immunology, Geron Corporation, Menlo Park, CA
Title: Differentiation of Human Embryonic Stem Cells to Insulin Producing Cell Clusters

Manfred R. Koller, PhD.
President & CTO, Cyntellect Inc. San Diego, CA
Title: Automated high-throughput cell imaging coupled with in situ laser-mediated cell manipulation on LEAP

Wen Tsang, PhD.
Senior Vice President, R&D, AmCyte, Inc. Santa Monica, CA
Title: Development of Microencapsulated Islets for the Treatment of Diabetesitle

Tim Allsopp, PhD.
Chief Scientific Officer, Stem Cell Sciences UK Ltd., Edinburgh, EH 9 3JQ UK
Title: Stem Cell Scale-up & -out: mind your Ps and Qs

Dr. Eugen Kolossov
Senior Scientist, Axiogenesis AG, Köln, Germany
Title: Lineage selection from differentiating transgenic ES cell culture: route to tissue engineering and teratoma free cell replacement therapy

Ulrich Hoffmueller, PhD., MBA
Chief Business Officer, Epiontis GmbH, Berlin Germany
Title: DNA Methylation Analysis-A Novel Approach for Cell Characterization in Regenerative Medicine

Peter Sartipy, Ph.D.
Chief Operating Officer, Cellartis AB, GÖTEBORG Sweden
Title: The potential and use of human embryonic stem cells in drug discovery

Donald J. Brown
Chief Executive Officer, Arteriocyte, Inc., Cleveland, OH

Ann Tsukamoto, PhD.
Vice President of R&D, StemCells, Inc., Palo Alto, CA
Title: Developing Human Neural Stem Cells for Clinical Applications

Howard Y. Chang, MD, PhD.
Assistant Professor, Program in Epithelial Biology, Stanford University Medical School, Stanford, CA
Title: Stemness redux: Shared and specific gene expression programs in stem cells and cancer

Gregory A. Bonfilio
Anthem Venture Partners, Santa Monica, CA

Madhusudan V. Peshwa, Ph.D.
Vice President, Research and Development, Maxcyte, Inc., Gaithersburg, MD
Title: Overcoming enablement challenges in the development and manufacture of therapeutic stem cell products

Jennie P. Mather, PhD.
Founder, President & Chief Scientific Officer, Raven Biotechnologies, Inc. South San Francisco, CA
Title: The Use of Fetal Tissue Stem or Progenitor Cell Lines as Drug Discovery Tools

Kitipan V. Arom, MD.
Bangkok Heart Hospital, Bangkok, Thailand
Title: Thoracoscopic intramyocardial autologous angiogenic progenitor cell (APC) transplantation for cardiomyopathy

Lola M Reid, PhD.
Departments of Cell and Molecular Physiology & Biomedical Engineering, University of North Carolina, School of Medicine, Chapel Hill, NC
Title: The Human Liver Stem Cell Compartment

Special Bioethics lecture by Professor William Hurlbut, MD. Consulting Professor & Member of the US President's Bioethics Committee

Panel Discussion on January 25, 2006 with experts from:

Law firm: Stacy Taylor, Partner Life Sciences Industry Team, Foley & Lardner LLP, San Diego
Non-Profit: Kenneth R. Rogulski, PhD. Scientist, Intellectual Property Management Dept. West Virginia High Technology Consortium Foundation, Wheeling, WV
Academia: William B. Hurlbut, MD. Stanford University Medical School, Stanford, CA
Biotech Industry: Jennie P. Mather, PhD. President, Raven Biotechnologies, Inc., South San Francisco, CA
Venture Capital: Gregory A. Bonfiglio, Anthem Venture Partners, Santa Monica, CA
Science Jounalist: TBA
Business Jounalist: TBA

Key Sessions
The most-up-to-date developments will be addressed:
Genetic and Transcriptional Controls of Stem Cells
Control of Cell Differentiation and Reprogramming
Self-renewal and Maintenance of Pluripotency
Human and Cancer Stem cells & their Propagation
Mutational analysis & Epigenetic Regulation
Applications of Stem Cells in various Human Diseases
Tissue Organ culture and Transplantation
Biopolymer applications in Biomedical Engineering
Stem Cells towards Developing Therapeutics

ABSTRACTS

Developing Human Embryonic Stem Cell-based Therapy for Parkinson’s disease
Ana Sofia Correia, PhD student (from Professor Patrik Brundin’s Lab)
Faculty of Medicine, Lund University, Lund, Sweden

A. S. Correia1, A. Brederlau2, S. Anisimov1, M. Elmi2, G. Paul1, L. Roybon1, P. Brundin1, P. S. Eriksson3 and J.-Y. Li1
1. Neuronal Survival Unit, Wallenberg Neuroscience Center, Dept. of Experimental Medical Science, Lund University, BMC A10, 221 84 Lund, Sweden
2 Institute of Anatomy and Cell Biology, Göteborg University, Box 420, SE-405 30 Gothenburg, Sweden
3 The Arvid Carlsson Institute for Neuroscience, Institute of Clinical Neuroscience, Sahlgrenska University Hospital, Göteborg University, SE-413 45 Gothenburg, Sweden.

Human embryonic stem cells (hESCs) may be a source of dopamine (DA) neurons for cell replacement in Parkinson’s disease. Transplantation success is limited by teratoma formation and by low cell survival or loss of DA phenotype. We have found that hESCs must be differentiated extensively in culture to avoid teratoma formation after grafting. After increasing time in culture, however, neurons are less likely to survive intracerebral implantation. We aim to eliminate residual undifferentiated hESCs and maintain a high survival rate of grafted neurons. We also study DA neuron survival and phenotype stability after genetic modification and addition of trophic factors.

Development of Pluripotent Stem Cell Therapies for Ischemic Injury and Disease
Robert J Deans, PhD. Vice President of Regenerative Medicine, Athersys, Inc., Cleveland, OH

Pluripotent stem cell cultures, such as the MAPC (Multipotent Adult Progenitor Cell), hold much promise for cell therapeutics based on their broad range of tissue regeneration and low immunogenic threshold. This is further enabled by the ability to expand cytogenetically and biologically stable stem cells to many clinical doses, providing consistent cell products for clinical use. Pluripotent stem cells have shown physiological benefit in pre-clinical models of ischemic injury to the heart, brain, and peripheral vasculature. In both the rat and pig, regional delivery of pluripotent stem cells to the peri-infarct zone in acute myocardial infarct results in significant improvement in wall function over 4 to 6 weeks of study. In rat models of peripheral vascular disease, significant increases in capillary density and retention of cells at vascular sites have been demonstrated. Using both allogeneic and xenogeneic stem cell treatments in rat stroke or ischemic injury models, strong performance improvements are correlated with retention of cells and localization throughout the brain. Clinical evaluation of pluripotent stem cells in ischemic injury provides safety with a high probability of benefit, and stages use of these cells for chronic degenerative diseases and tissue regeneration

Billions of embryonic stem cells for cytotherapy: quality controlled expansion in bioreactors
Nicole I. zur Nieden, PhD. Post doctoral Fellow in the lab of Dr. Derrick E. Rancourt
Institute of Maternal & Child Health, University of Calgary, Calgary, AB, CANADA

Nicole I. zur Nieden1, Jaymi T. Cormier2, Michael S. Kallos2, Derrick E. Rancourt1
1Institute for Maternal & Child Health, University of Calgary, 3330 Hospital Drive NW, Calgary, AB T2N 4N1, 2Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4

With the recent emergence of tissue engineering, a field which offers promising treatment alternatives for numerous degenerative diseases, a greater interest has been drawn towards the potential use of pluripotent embryonic stem cells (ESCs) as the basic material for tissue regeneration. For clinical application of this technology, a quality controlled, reproducible culture system is necessary for the expansion and differentiation of ESCs. The current study describes a successful protocol for the expansion of murine ESCs as aggregates in suspension bioreactors. In this system, excessive agglomeration of the cells was controlled through the manipulation of hydrodynamic shear and inoculation density.

Development of Microencapsulated Islets for the Treatment of Diabetes
Wen Tsang, PhD.
Senior VP R&D, AmCyte, Inc. Santa Monica, CA

Cellular replacement therapy represents a promising strategy for treatment of diabetes. However, such an approach is limited by the inadequate availability of human donor tissue. We have developed culture processes to promote intermediate progenitor growth and generate functional islet-like aggregates from adult human pancreatic tissue. These regenerated islets secrete insulin in response to glucose stimulation as well as lower blood glucose and secrete c-peptide in diabetic animals. These processes serve as platforms for further scale up studies to produce large amount regenerated islets for transplant as well as to elucidate cell proliferation and differentiation mechanisms of human pancreatic endocrine cells in vitro.

Automated high-throughput cell imaging coupled with in situ laser-mediated cell manipulation on LEAP
Fred Koller, Ph.D., President and CTO, Cyntellect, San Diego, CA.

LEAP™, based on robust semiconductor manufacturing technologies, enables in situ manipulation of cell types that are refractory to conventional approaches (e.g., FACS, electroporation, etc.). High-throughput imaging is used to identify cells based on multiple morphologic and fluorescence parameters. Selected cells are then automatically laser-irradiated to achieve cell purification (with high-yield and purity) and/or transfection (with high-efficiency and low-toxicity) at rates exceeding 1,000 per second. LEAP has been used for efficient purification of valuable cells in samples ranging from 101-108 cells, and high-efficiency transfection of difficult human cell types with a wide range of compounds and macromolecules. These capabilities of LEAP, combined with its closed-system sterile processing, will find use in various stem cell research and clinical applications.

Adult Neural Progenitor Cells and Cell Replacement Therapy for Huntington’s Disease
Bronwen Connor, PhD. Department of Pharmacology and Clinical Pharmacology, Faculty of Medical and Health Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand

The presence of progenitor cells in the adult mammalian brain raises the exciting possibility that adult neural progenitor cells could be used therapeutically for repair of neuronal loss associated with neurodegenerative diseases such as Huntington’s disease (HD). Potential therapeutic uses of adult neural progenitor cells include transplantation to replace lost cells and the activation of endogenous progenitor cells to provide “self-repair.” In order for endogenous progenitor cells to be useful therapeutically, methods need to be developed to direct progenitor cells to migrate and generate new neurons in specific areas of neuronal cell loss. Using in vivo gene transfer, we have demonstrated the role Brain Derived Neurotrophic Factor (BDNF) plays in regulating and directing the proliferation, migration and differentiation of endogenous progenitor cells in both the normal brain and in the quinolinic acid (QA) lesion model of HD. We also examined the use of adult neural progenitor cells for cell transplantation therapy for the treatment of HD. The results of this study demonstrate that adult neural progenitor cells survive transplantation into the striatum of the QA lesion model of HD and generate new striatal neurons, resulting in a significant reduction of the motor function impairment observed in HD. The results of these studies demonstrate the potential use of adult neural progenitor cells for cell replacement therapy and aids in the development of novel therapeutic strategies for the treatment of HD. These projects were supported by the Neurological Foundation of NZ and the Marsden Fund, Royal Society of NZ.

The Potential of Cord Blood Stem Cells for Use in Tissue Engineering of the Eye
David T. Harris, PhD. Professor, Department of Microbiology and Immunology &
Director, Stem Cell Bank, University of Arizona, Tucson, AZ

David T. Harris1, Xianghui He2, Daniel Camacho2, Veronica Gonzalez2 and John C. Nichols3
1University of Arizona and CBR Systems, 2Dept. Immunobiology and 3Dept. Ophthalmology; University of Arizona, Tucson, AZ 85724

Cord blood (CB) is a rich source of hematopoietic stem cells (SC). These SC are neonatal in origin and may be an alternative to the use of fetal SC for many applications. In this study we examined the plasticity of CBSC for use in tissue engineering. CBSC were observed to differentiate into an epithelial phenotype in a corneal epithelial culture system. The resultant cell sheet was morphologically indistinguishable from corneal epithelial cells, expressed the corneal epithelial specific cytokeratin k3, and could be transplanted into large animals. The first animal demonstrated a clear cornea with a minimal degree of scarring and neovascularization. Epithelial cells resurfaced the cornea to include a basal layer covered by four to five layers of stratified squamous cells, although it was thinned in some regions to about two to three cell layers. A few rare goblet cells were observed within the epithelium. The other animal showed similar results, although some stromal scarring and neovascularization were evident. In some regions the epithelium appeared to be poorly organized. Rare goblet cells were observed. Areas of sparse inflammatory cells included mostly neutrophils and a few atypical inflammatory cells. Additional animals are currently being transplanted and evaluated. Human cord blood stem cells could proliferate to form tissue grafts in a culture system optimized for corneal epithelium. Transplantation of cultured grafts onto a rabbit cornea with limbal stem cell deficiency can form an epithelium that is optically clear. CD34-depleted and CD34-enriched cord blood stem cells showed the greatest potential to maintain a clear cornea with the least vascularization and scarring. Overall, CBSC appear capable of differentiating into nervous, endothelial and epithelial tissues under proper culture conditions, and demonstrate potential for the treatment of limbal stem cell deficiency. In an effort to identify molecular pathways involved in stem cell differentiation, we compared CB CD34+CD133+ cells with their progenies using a cDNA microarray containing 22,000 human cDNA clones. A total of 139 genes were differentially expressed between CBSC and their progenies. Among the genes showing the greatest differential expression levels in stem cells were: psoriasin 1, CRHBP, HDAC3, MLLT3, HBEX2, SPINK2, c-kit, H2BFQ, CD133, HHEX, TCF4, ALDH1A1 and FHL1. Thus, CBSC appear to be highly plastic in their ability to differentiate into other tissue types and may be comparable to fetal SC for many tissue engineering applications.

How Will Intellectual Property Factors Affect the Commercialization of Stem Cell Therapeutics?
Todd N. Spalding, Associate, Life Sciences Industry, Foley & Lardner LLP., Washington, D.C.

With the rest of the world, the US is struggling to accommodate scientific, ethical, business, and political considerations that will determine when or even whether different stem cell innovations realize their full potential. These considerations are conditioned by intellectual property factors, the effects of which recall but also diverge from past experience with paradigm-breaking technologies. This presentation reviews the current patent situation globally for stem cell-related developments and reflects on whether this nascent field of regenerative medicine will be advanced or hindered by IP factors.

Foley & Lardner LLP is a full-service law firm, providing an integration of services in business law, regulatory and public affairs, intellectual property, health law, financing, real estate development, and taxation. Our attorneys work closely with universities, research facilities, and public and private companies to develop comprehensive policies covering issues at the core of stem cell technologies. At Foley, we understand that the challenges and complexities of stem cell research require a complete and thorough approach. We work with you to meet head-on the challenges that you face, with thoughtful solutions not found elsewhere. Please visit us online at

Lineage selection from differentiating transgenic ES cell culture: a route to tissue engineering and teratoma free cell replacement therapy
Eugen Kolossov, PhD. Senior Scientist, Axiogenesis AG, Köln, Germany

Relatively low content of differentiated cells of certain tissue type as well as contamination with tumorogenic undifferentiated ES cells are major challenges to the ES cell based cell replacement therapy. We show that lineage selection from differentiating transgenic ES cell culture based on the stable tissue specific expression of both drug resistance and reporter cassettes provides both high purity and high yield of the selected differentiated cells. Parallel selection of two and more cell types could lead to formation of tissue-like structure. Transplantation of the purified cells results in a high grafting efficiency along with negligible level of teratoma formation

Advances in Cardiac Stem Cell Therapy
Yerem Yeghiazarians, MD. Assistant Professor of Medicine & Director of Cardiac Stem Cell Translational Development Program, Division of Cardiology, University of California at San Francisco Medical Center, San Francisco, CA

The adult heart muscle cells (cardiomyocytes) are believed to be mature, terminally differentiated cells that cannot regenerate new cells. After a heart attack, irreversible loss of contracting heart muscle cells occurs. Current therapies designed to treat heart attack patients in the acute setting aim to open the blocked coronary arteries. Unfortunately, it is rarely possible to rescue the at-risk heart muscle cells from some degree of irreversible injury and death. Experimental studies in both animals and humans have revealed encouraging results when stem cells are injected into the heart in the areas of myocardial infarction. These therapies appear to result in improvement in the contractile function of the heart.

Differentiation of Human Embryonic Stem Cells to Insulin Producing Cell Clusters
Anish S. Majumdar, Ph.D. Senior Director, Immunology, Geron Corporation, Menlo Park, CA

Recent success in cadaveric islet transplantation for patients with type I diabetes has increased interest in discovering an alternative source of stem cells with potential to differentiate into pancreatic ? cells. Human embryonic stem cells (hESCs), which are immortal and capable of both self-renewal and differentiation, could be used for this purpose. We have developed a three-stage differentiation protocol in which human embryonic stem cells (hESCs) are allowed to differentiate to definitive endoderm. In the presence of selective growth factors, extracellular matrix and maturation inducing factors, the pancreatic component of the endoderm differentiates into insulin and glucagon expressing islet-like cell clusters. These clusters, similar to human islets, secrete C-peptide in response to glucose challenge. In vitro and in vivo properties of these islet-like cells derived from hESCs will be discussed at the meeting.

Assessing the Genetic and Epigenetic Stability of Stem Cells
Evan Snyder, MD., PhD. Professor and Director of Program in Developmental & Regenerative Cell Biology, The Burnham Institute for Medical Research, Neuroscience & Aging Research Center (Stem Cell Research Center), La Jolla, CA

Searching for “stemness” genes has suggested that gene expression alone is insufficient to define plasticity and lineage-specification. We hypothesized that stem cell behavior should also be characterized by epigenetic processes, specifically chromatin state which in turn determines the degree of transcriptional activity. We have examined the dynamics of epigenetic chromatin modification first in a “pluripotent” human ”embryonic” stem cell (hESC) and then in a “multipotent” human “somatic” stem cell (i.e. germ-layer-restricted). The latter was represented by the human neural stem cell (hNSC). hNSC were derived either in vitro from hESCs (“secondary”) or isolated directly from the human fetal neuroectoderm (“primary”), allowing these two derivations also to be compared via epigenetic marks and the stability of their commitment. We demonstrate that analysis of the dynamic transition between distinct chromatin states in human stem cells provides evidence for, and offers one mechanism subserving, epigenetic control of stem cell behavior.

Stemness redux: Shared and specific gene expression programs in stem cells and cancer
Howard Y. Chang, M.D., Ph.D. Program in Epithelial Biology, Stanford University School of Medicine, Stanford, CA

Stem cells from different lineages share the capacity for self-renewal and pluripotency, but whether a common molecular program underlies these functions is controversial. Here we analyze the global gene expression patterns from 95 human and 75 mouse microarrays of stem cells and their differentiated counterparts at the level of gene modules, groups of genes with shared biological function or regulation. We describe each stem cell by a combination of activated and deactivated gene modules, and identify gene modules associated with self-renewal and pluripotency. These results suggest a new classification scheme for stem cells and identify shared transcriptional programs between stem cells and cancer.

Role of microRNAs during cardiac progenitor differentiation and cardiogenesis
Deepak Srivastava, MD. Gladstone Institute of Cardiovascular Disease, University of California San Francisco, San Francisco, CA

Gradients of signaling and transcription factors result in distinct cellular responses during organ formation, suggesting that the precise dose of major regulatory proteins must be tightly controlled. MicroRNAs are phylogenetically conserved small RNAs that regulate translation or stability of target messenger RNAs, providing a mechanism for protein dose regulation. MicroRNA 1-1 (miR-1-1) and miR-1-2 are specifically expressed in cardiac and skeletal muscle progenitor cells during development and are transcriptionally regulated by central myogenic factors, including serum response factor, MyoD and Mef2. Gain-of-function of miR-1 in the developing mouse heart led to a decreased pool of proliferating ventricular cardiomyocytes and early exit from the cell cycle. In contrast, loss of miR-1 in Drosophila led to a defect in cardiac differentiation, lineage determination and maintainance of cardiac-specific gene expression. To identify the transcripts affected by miR-1 in vivo, we developed a novel algorithm for microRNA target identification, incorporating features of RNA structure and target accessibility. In mice, we found that mRNA encoding endogenous Hand2, a cardiac transcription factor that promotes ventricular cardiomyocyte expansion, is a target of miR-1. In flies, miR-1 targets the Delta transcript, which encodes a ligand for the Notch receptor. Notch signaling is involved in cardiac lineage decisions and differentation, providing a potential mechanism for the observed progenitor cell defect. This work reveals an evolutionarily conserved mechanism for regulation of the heart and suggests that miR-1s titrate the effects of critical cardiac signaling and transcriptional pathways to control cell fate, differentiation, and proliferation during cardiogenesis.

Human Embryonic Stem Cells in Drug Discovery and Toxicology
Peter Sartipy, Ph.D. Project Manager, Cellartis AB, GÖTEBORG Sweden

The use of human embryonic stem cells (hESC) in drug discovery spans from early target finding and evaluation studies, via the use of functional cells in screening and metabolism studies, to the use of various stem cell technologies in toxicity testing. In addition, human stem cells provide a platform for the development of drugs that may activate and mobilize endogenous pools of stem cells. Cellartis AB is focusing on development of hESC derived functional human cell types, e.g. hepatocytes and cardiomyocytes, and to use the hESC platform for the development of novel toxicity tests.

Somatic Cell Cloning and Nuclear Reprogramming via Cloning from de differentiation to re-differentiation – State of the Technology and Future Applications
Xiangzhang (Jerry) Yang, PhD. Center for Regenerative Biology, University of Connecticut, Storrs, CT

Cloning whole animals from adult somatic cells has been succeeded in various mammalian species. However, the overall efficiency has been very low. Therefore, an active area of research on somatic cloning is to improve the cloning efficiency to a commercially feasible level. State of the technology and biological factors affecting the efficiency of the technology will be reviewed. In this presentation, I will present our research on animal cloning in cattle, pigs, rabbits as well as mice, the health and safety of cloned animals and our approach to study nuclear reprogramming via candidate and global gene analyses. I will discuss the State of Connecticut stem cell bill to legally and finally support human somatic cell nuclear transfer and stem cell research, the status of our team research in this exciting area and my dreams for potential applications of somatic cell nuclear transfer and embryonic stem cells in agriculture, basic biological research, and potential biomedicine therapy in the next few years.

Tissue Engineering Potential of Embryonic Stem Cells Using Three-Dimensional
Biodegradable Scaffolds
G. Rasul Chaudhry, PhD. Department of Biological Sciences, Oakland University, Rochester, MI

G. Rasul Chaudhry1, Donggang Yao2, Miguelangelo PerezCruet3, and David Svinarich3 1Department of Biological Sciences, Oakland University, Rochester, MI, 2School of Polymer, Textile & Fiber Engineering, Georgia Institute of Technology, Atlanta, GA, 3Providence Medical Center, Southfield, MI

There is a great deal of interest in developing strategies for regenerating, repairing and replacing damaged or lost tissues and organs due to debilitating diseases and traumatic injuries. Embryonic stem cells (ESCs) have the ability to give rise to any cell type and thus have unlimited potential for therapeutic applications. Our working hypothesis is that ESC-derived progenitors of specific lineages, cultivated in biodegradable scaffolds, will mimic in vivo 3-dimensional growth to yield suitable grafts for transplantation. To study the ex vivo growth potential of ESC-derived progenitors, they were seeded into biodegradable scaffolds that were fabricated using polycaprolactone and poly ethylene oxide. The results of in vitro experiments showed that the ESC-derived cells colonized the scaffolds and migrated into the porous scaffold structure. The in vivo studies showed, while the ESCs injections developed into teratomas, the scaffold seeded ESCs and their derivatives had restrained cell growth and yielded desired tissues.

DNA Methylation Analysis - A Novel Approach for Cell Characterization in Regenerative Medicine
Ulrich Hoffmueller, PhD, MBA, Chief Business Officer, Epiontis GmbH, Berlin, Germany

The epigenetic phenomenon of DNA methylation possesses a large potential for cell characterization. Specific methylation patterns correlate with differentiation processes, cell type and long-term cell functions. In this talk it will be demonstrated how DNA methylation analysis can be applied for quality control in regenerative medicine. In a first step, using genome wide discovery methods, DNA methylation candidate markers are identified. Upon marker validation they can be applied to determine identity, purity and potency of cell samples. Several high performance methylation assay methods permit sensitive and quantitative characterization of cellular therapeutics. Applicability for research and routine use will be discussed.

Stem Cell Scale-up & -out: mind your Ps & Qs.
Timothy E. Allsopp, PhD. Chief Scientific Officer, Stem Cell Sciences UK Ltd, West Mains Rd, Edinburgh EH9 3JQ UK

Stem cells are a cellular resource providing unprecedented potential for biological discovery & future medicines. Significant challenges are presented to researchers seeking to provide stem cells of a quantity & quality for these applications. For example defining appropriate expansion conditions to maintain indefinite self-renewal, designing animal component free culture and the development of a production process with minimal cell manipulation. Human stem cells are difficult to control in culture with many cell types requiring convoluted systems to maintain indefinite expansion without unintentional differentiation. A summary will be presented on the latest efforts within the bio-industry of expanding human stem cells.

Overcoming enablement challenges in the development and manufacture of therapeutic stem cell products
Madhusudan V. Peshwa, Ph.D. Vice President, Research & Development, Maxcyte, Inc., Gaithersburg, MD.

The ability to enhance biological function and control cell fate decisions in a transient manner represents a paradigm shift in the development of cellular therapeutics. While new learnings are continuing to evolve from our understanding of molecular pathways involved in developmental biology, their utilization in the manufacture & characterization of effective cellular therapeutic products for regenerative medicine applications has not been addressed. MaxCyte is a clinical stage biotechnology company developing cellular therapeutics utilizing its proprietary cell-loading technologies. Cell-loading technologies are fundamental to cell-based therapeutics, gene therapy, and many biopharmaceutical manufacturing applications. MaxCyte has developed an efficient, non-viral, scaleable, GMP-compliant, functionally closed system for manipulation of cellular function. The company’s pipeline includes an engineered tumor vaccine, currently under investigation in a Phase I/II clinical study, for the treatment of B-cell Chronic Lymphocytic Leukemia (B-CLL) and pre-clinical candidates targeting applications in oncology, infectious diseases and regenerative medicine. In addition to these therapeutic programs, MaxCyte has in specific instances selectively licensed its technology to enable development of ex vivo cell therapies and for manufacture of biopharmaceutical products being developed by other commercial entities. Current partners are working on development of therapeutics for pulmonary disease, infectious diseases, oncology, and regenerative medicine applications. The ability to manipulate cellular function in a safe, reliable, consistent and scalable manner represents a significant milestone for engineering the potency of cellular products and in enabling manufacture of stem cell based therapeutics.

The Use of Fetal Tissue Stem or Progenitor Cell Lines as Drug Discovery Tools
Jennie P. Mather, PhD. Founder, President & Chief Scientific Officer, Raven Biotechnologies, Inc. South San Francisco, CA

Jennie P. Mather, Penny Roberts, Deryk Loo, Suzanne Coberly, Raven Biotechnologies, Inc. South San Francisco, CA

As an embryo develops, the differentiation potential of cells in each fetal organ becomes progressively more limited until most cell types are terminally differentiated in the adult. Fetal tissue stem cells can then be defined as those cells that have the capacity to divide indefinitely and can differentiate into one or more terminally differentiated cell types. The use of defined serum-free media has allowed the establishment of a number of human fetal tissue stem cell lines arrested in specific stages of the differentiation process but capable of indefinite expansion in vitro while in this arrested state. These lines have been used to raise monoclonal antibodies to their distinctive cell surface proteins. Screening these Mabs against tumor tissue has provided a rich source of therapeutic antibodies to both known and novel onco-fetal antigens.

Thoracoscopic intramyocardial autologous angiogenic progenitor cell (APC) transplantation for cardiomyopathy
Kitipan V. Arom, MD. Bangkok Heart Hospital, Bangkok, Thailand

Kitipan V. Arom, MD., P Ruengsakulrach, MD., Vibul Jotisakulratana, MD., Amit Patel, MD. Bangkok Heart Hospital, Bangkok, Thailand

Background: Stem cell therapy is rapidly emerged as a potential technology for the treatment of cardiomyopathy. Cells can be delivered directly into myocardium with minimally invasive technique. The objectives are to assess the safety/ efficacy and short term results of this approach.

Methods: Between May and December 2005, 26 patients underwent thoracoscopic APCs injection. The mean age was 57.6 ± 12.4 years. Twenty four (92.3%) were male. Eight had dilated cardiomyopathy (DCM) and eighteen had ischemic cardiomyopathy (ICM). In ICM group, six patients had previous PCI, eight had previous CABG. Mean ejection fractions were 34.2 ± 17.7 % in the ICM and 27.0 ± 9.8 % in the DCM. Twenty one patients (80.8%) were in NYHA Class III & IV. The cells were derived and expanded from the autologous blood, separated mainly CD34+ cells. The number of cells prior to injection was 27.2 ± 21.4 x 106 cells (viability was 92.6 ± 4.3 %). In the ICM, twelve had cell injection and six had cell injection plus OPCAB. The cells were injected into the nonviable myocardium assessed by cardiac MRI (CMR). In DCM, all eight had cells injection in entire wall of the left ventricle. All patients had preoperative characteristics, intraoperative variables and postoperative results recorded including major advance cardiac events (MACE).

Results: At two months, there was no MACE. There was one death on the third postoperative day after redo CABG and cell injection, most likely from pulmonary emboli. Six-minute walk test improved from an average of less than 300 meters to more than 400 meters. Majority of functional class improved from class III to II. Six patients had follow-up CMR showed decrease in size of the scar and improve in wall thickness (figure). The ejection fraction improved from 30.4 to 45%.

Conclusions: Thoracoscopic intramyocardial autologous APCs transplantation is feasible and safe in both DCM and ICM patients. The early results are good with phase two trial and the long term results are in progress. The technique is slightly more invasive than trans-catheter approach but provides better target for cell injection.


The Human Liver Stem Cell Compartment
Lola M Reid, PhD. Departments of Cell and Molecular Physiology & Biomedical Engineering, University of North Carolina, School of Medicine, Chapel Hill, NC

Eva Schmelzer1, Hiroshi Kubota1, Lili Zhang1, Randall McClelland1, Alaa Melhem1, Hsin-lei Yao2, Eliane Wauthier1, Huifei Liu1, William Turner2, Nick Moss1, John Ludlow4, Andrew Bruce4, Sonya Sherwood4, Megan Christie4, Mark Furth4, and Lola M Reid1-3 . 1Departments of Cell and Molecular Physiology and 2Biomedical Engineering, UNC School of Medicine, Chapel Hill, NC 27599; 4Vesta Therapeutics, Durham, NC 27713.

Human liver stem cell compartments are comprised of hepatic stem cells (HpSCs), hepatoblasts (HBs), unipotent hepatic progenitors, and mesenchymal progenitors (angioblasts and stellate cell precursors), located in limiting plates in fetal livers and Canals of Hering in postnatal livers. The antigenic profiles and ex vivo clonogenic expansion conditions for each subpopulation have been defined. HpSCs are found at 0.1-0.7% of the cells in livers of all age donors, are 7-10 mm in diameter, form morphologically uniform colonies, and express EpCAM and an array of hepatocytic, biliary and stem cell markers but no hemopoietic markers, mesenchymal cell markers, or alpha-fetoprotein (AFP). The HpSCs give rise to HBs, that are larger (10-12 mm), with overlapping antigenic and biochemical profiles to HSCs but differ in expression of ICAM1, P450-3A7, AFP, and yield cord-like colonies with canaliculi. The HBs dwindle in numbers with donor age remaining as cell aggregates tethered to the ends of Canals of Hering. Enrichment for hepatic progenitors is achieved by immunoselection for EpCAM+ cells. Self-replication at ~one division/day and with high telomerase levels is observed under specific culture conditions. EpCAM+ cells form liver tissue when transplanted into SCID/nod mice. Potential medical applications include the treatment of liver disease by cell therapies such as cell transplantation and/or bioartificial livers. Funding was provided by Vesta Therapeutics, by NIH grants (DK52851, AA014243, IP30-DK065933) and by a Department of Energy Grant (DE-FG02-02ER-63477).

Developing Human Neural Stem Cells for Clinical Applications
Ann Tsukamoto, Ph.D. VP, Research & Development, StemCells, Inc., Palo Alto, CA

StemCells, Inc. has purified and expanded tissue derived human neural stem cells (hCNS-SC). Preclinical mouse studies, in a non-degenerative environment show that the human cells engraft reproducibly, populate the endogenous stem cells niche and send progeny to the various parts of the brain where they mature into the 3 major cells types of the CNS, neurons, astrocytes and myelin producing oligodendrocytes in a site-appropriate manner. The hCNS-SC have also been transplanted into mouse models of diseases or injuries. In the mouse model, that recapitulates the human lysosomal storage disorder, infantile NCL (INCL), we have shown that the hCNS-SC can be used as a cellular delivery vehicle to deliver missing enzyme across the blood-brain barrier. Moreover, enzyme production is sustained throughout the lifespan of the mouse. We have shown that these hCNS-SCs produce other housekeeping lysosomal enzymes as well and could potentially be used to treat the spectrum of LSDs affecting the CNS, where a secreted enzyme is missing or defective. In the shiverer mouse, deficient in myelin production, hCNS-SC produce myelin from new oligodendrocytes. Myelin production is also observed in mice receiving a spinal cord contusion injury and could be one mechanism whereby the hCNS-SC are able to partially restore motor function in this injury model. Remyelination by human neural stem cells could also lead to neuroprotection in disorders such as cerebral palsy and multiple sclerosis and thereby provide a therapeutic benefit. These hCNS-SC have been established into qualified master and working cell banks for therapeutic application into patients with INCL or Late-INCL.

Hematopoietic Stem Cells: From Bench to Bedside
Daniel Kraft, MD. Fellow, Hematology/Oncology/BMT, Stanford Univ. School of Medicine, Stanford, CA

Stem cell therapy has been underway for over 30 years in the field of bone marrow transplantation. This talk will be focused on the biology of hematopoietic stem cells and their current and evolving use in the clinic for the treatment of malignancy and a widening array of non-malignant disorders. Developments in the push for therapeutic applications utilizing bone marrow stem cells for use in the rapidly evolving field of regenerative medicine will also be covered.

Stem Cells, Embryos and Ethics: Is There A Way Forward?
William B. Hurlbut, MD. Consulting Professor & US President’s Bioethics Committee
Stanford University Medical School, Neurology and Neurological Sciences, Stanford, CA

The controversy over embryonic stem cell research has generated a global debate that places the positive prospect of biomedical advance in conflict with traditional moral values. In this session, we will explore the fundamental sources of the moral impasse and discuss the possibility of a technological solution that can sustain social consensus for progress in stem cell biology. Drawing on the President's Council on Bioethics report "Alternative Sources of Human Pluripotent Stem Cells," we will focus on Altered Nuclear Transfer, a proposal that has recently been the subject of proof of principle research by MIT biologist Rudolf Jaenisch.