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Day Two: Stem Cell Sources, Culture and Expansion


TUESDAY, AUGUST 30

8:30 Morning Coffee

8:45 Chair's Remarks
Dr. Anthony Johnson, Business Area Manager, Invitrogen

keynote presentation
8:50 Stem Cells in Regenerative Medicine and Reproductive Biology 
Dr. George Q. Daley, Associate Professor of Pediatrics, Children's Hospital, Boston
This presentation will highlight the use of embryonic stem cells to unravel the molecular mechanisms of blood and germ cell formation, and the use of somatic cell nuclear transfer to create customized stem cell lines for treatment of genetic disease.


Cell Culture

9:35 Defining the Feeder Activities for Human ES Cell Self-renewal
Dr. Ren-He Xu, Senior Scientist, Research and Development, WiCell Research Institute 
Current culture conditions for human embryonic stem cells (hESCs) require unidentified molecules from mouse or human feeder cells, potentially transmitting xenogeneic or allogeneic pathogens. We demonstrate that hESCs are subjected to high levels of bone morphogenetic protein (BMP) activity when cultured without support of the feeder cells. The BMP antagonist noggin synergizes with basic fibroblast growth factor to repress BMP signaling and sustain undifferentiated proliferation of hESCs in the absence of feeder cells or feeder-conditioned medium.

10:05 Stem Cell Differentiation using Combinatorial Cell Culture
Dr. Yen Choo, CEO, Plasticell Limited 
Plasticell is a biotechnology company that has developed a novel cell culture method 'Combinatorial Cell Culture' which allows it to assay large numbers (up to millions) of different cell culture protocols in parallel, to derive new methods of growing or differentiating cells. Combinatorial Cell Culture will have widespread utility throughout cell biology, and particularly in the differentiation of stem cells. There the method has been used to study the behavior of pluripotent embryonic stem cells under large numbers of culture conditions, generating new protocols for the directed differentiation of these cells into interesting lineages. Plasticell is focused on identifying and dissecting pathways of cell differentiation with the aim of generating regenerative medicines. 

10:35 Coffee Break, Poster and Exhibit Viewing

11:15 New Technologies to Advance Adult Stem Cell Applications
Dr. James L. Sherley, Associate Professor, Div. of Biological Engineering, Massachusetts Institute of Technology 
Among the barriers to developing adult stem cell-based applications for biomedicine and biotechnology are the difficulty in expanding them in culture, and the lack of molecular markers that uniquely identify them. We have focused on a unique property of adult stem cells, asymmetric self-renewal, as the basis for developing new technologies for their expansion and identification. Proof of principle has been established in rodent tissue models, and current studies evaluate the application of these technologies to two different human adult stem cell types, liver and blood.

11:45 Interactive Panel Discussion with Morning Speakers

12:15 Luncheon Technology Workshop  Sponsored by 
Human Embryonic Stem Cells: Derivation, Expansion, and Characterization
Dr. Ian Lyons, Research Area Manager, Stem Cells, Invitrogen
In the field of stem cell biology there remains fundamental progress to be made in the development of methods for the propagation and maintenance of HESCs in vitro, preferably using defined components not of animal origin. Developing defined culture conditions and defined stem cell populations, to help the field of stem cell therapies be able to make more rapid progress.

Stem Cell Sources

1:30 Chair's Remarks
Dr. John D. McNeish, Senior Director, Genetic Technology, Pfizer 
Global Research and Developmen

1:35 A Novel Approach to the Visualisation of Molecular Events
Sponsored by
Ms. Nancy Angus, Business Development, Sidec Technologies
SET visualizes the cellular events at a molecular level, in virtually any environment and biological context, thereby bridging the current gap between the test tube and human biology. Sidec Technologies AB was established year 2000, based on an innovation from the Karolinska Institute in Stockholm. Principal investors are the Karolinska Investment Fund, the Swedish Industrial Investment Fund and FEI company a leading international EM microscope manufacturer. Since 2004, Sidec has sales offices in Europe and in the US. 

1:50 Novel Products for ES Cell Research
Dr. Michelle Greene, Director, Stem Cell Segment, Chemicon International
As a leading provider of tools for the stem cell researcher, Chemicon is committed to advancing this important research area by meeting its unique needs. This talk will provide a brief overview of some of the novel cell lines, media, markers and kits we offer for stem cell research.

2:05 Science, Ethics and Therapeutic Potential of Human Stem Cells 
Dr. Stephen Minger, Stem Cell Biology Lab, Wolfson Centre for Age-Related Diseases, King's College London 
Human embryonic stem cells offer considerable promise in the treatment of significant human disease, but only if the requisite therapeutically important cell populations can be generated. Under license from the UK Human Fertilization and Embryology Authority, our lab has derived three human embryonic stem cell lines, including one that encodes the most common mutation for Cystic Fibrosis. Much of our current work is focused on the clinical translation of human ES cells for CNS, retinal, cardiac, endocrine and hepatic disorders.

2:35 Oct-4-Expressing Cells Isolated from Umbilical Cord Matrix are Multipotential Stem Cells
Dr. Kathy Mitchell, Assistant Professor, Pharmacology and Toxicology, University of Kansas 
We have isolated mesenchymal stem cells (MSC) from the human umbilical cord matrix (HUCM). These cells proliferate in culture and express markers found in other stem cells. Specifically, HUCM cells express the transcription factors Oct-4 and nanog, which are important for maintaining the undifferentiated, pluripotent state of embryonic stem (ES) cells. These Oct-4 expressing cells are found in the perivascular region of the umbilical cord. They can be differentiated into multiple cell types including neuronal, endothelial and epithelial cells and are therefore candidates for cell-based therapies. In contrast to ES cells, but like umbilical cord blood cells, HUCM cells display more immune tolerance and have been shown not to form tumors when injected in immune-compromised mice. Furthermore, these cells are easily accessible compared to bone marrow MSC, are more abundant than the MSC found in cord blood and they lack the ethical considerations of ES cells.

3:05 Hair Follicle Nestin-Expressing Stem Cells can Form Neurons
Dr. Robert M. Hoffman, Professor of Surgery, University of California-San Diego;
President, AntiCancer, Inc. 
We have recently shown the expression of nestin, the neural-stem-cell marker protein, is expressed in bulge-area stem cells of the hair-follicle. We used transgenic mice with green fluorescent protein expression driven by the nestin-regulatory-element (ND-GFP). The ND-GFP stem cells give rise to the outer-root sheath of the hair follicle as well as to a nestin-expressing interfollicular vascular network. In the present study, we demonstrate that ND-GFP-expressing stem cells isolated from the hair-follicle bulge area that are negative for the keratinocyte marker keratin 15 can differentiate into neurons, glia, keratinocytes, smooth muscle cells, and melanocytes in vitro. These pluripotent nestin-GFP-expressing stem cells are positive for the stem cell marker CD34, as well as keratin 15-negative, suggesting their relatively undifferentiated state. The apparent primitive state of the ND-GFP stem cells is compatible with their pluripotency. Furthermore, we show that ND-GFP-expressing stem cell-derived cells can differentiate into neurons after transplantation to the subcutis of nude mice. These results suggest that hair follicle bulge-area ND-GFP stem cells may provide an accessible, autologous source of undifferentiated multipotent stem cells for therapeutic application.

3:35 Refreshment Break, Poster and Exhibit Viewing

Sponsored by:


Labeling/Tracking Stem Cells

4:00 Label-Retaining Epithelial Cells in Mouse Mammary Gland Divide Asymmetrically Retaining Their Template DNA Strands
Dr. Gilbert Smith, Principal Investigator, MBTL,CCR, National Cancer Institute, NIH
An aspect of somatic stem cells that is indicative of their "stemness" is the characteristic of retaining radioactive nucleotides over long periods of time after labeling in vivo. This has been ascribed to their relative mitotic quiescence and/or to their very slow passage through the cell cycle. An alternative view was proposed by John Cairns in 1975. He posited that stem cells dividing asymmetrically in somatic tissues selectively retain their template DNA strands during cell division and partition the newly synthesized strands to their committed (and dispensable) daughters. In this way, somatic stem cells were protected from mutations resulting from errors in DNA replication. As an additional consequence of this, mechanism asymmetrically cycling stem cells would retain radio-nucleotide label received during their inception over long time periods. This characteristic of stem cells would also permit their relative insensitivity to protocols designed to kill cells through the incorporation of nucleoside analogs or senescence due to telomere shortening. Double labeling of long label-retaining mammary epithelial cells in situ provides direct confirmation of the existence of this mechanism for genome protection in the mouse mammary gland. The recent interest in tumor-initiating stem cells and their role in solid tumor maintenance, potentially through asymmetric cell division, marks this aspect of "stemness" as having increasing importance in the design of cancer therapeutics.

4:30 Tracking the Fate of Stem Cells by in vivo MRI
Dr. Joseph Frank, Chief, Experimental Neuroimaging Section LDRR, National Institutes of Health 
Magnetic labeling of cells provides the ability to monitor their temporal spatial migration in vivo MRI. Various methods have been used to magnetically label cells using coated superparamagnetic iron oxide (SPIO) nanoparticles. In this presentation, I will describe the different approaches used to label cells with contrast agents and show MRI and histologic results in various animal disease models. Magnetic Tagging of stem cells and other mammalian cells has the potential for guiding future cell-based therapies in humans and for the evaluation of cellular-based treatment effects in disease models.

5:00 Interactive Panel Discussion with Afternoon Speakers

5:30 Close of Day Two


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