ISSCR 2011International Society for Stem Cell Research

Thermo Scientific ISSCR 2011 Industry Symposium

Wednesday Industry SymposiaJune 15, 20118:30am - 12:30pm

ISSCR 2011 - Abstracts and BiographiesThermo Fisher Scientific - Industry Symposium

Plenary Lecture and Thermo Scientific StemCellExcellence Award winner – microRNA Screens: Optimization, Target Identification and Pathway Discovery in Stem Cell Reprogramming

Robert Judson, Blelloch Lab, USCF, Institute for Regenerative Medicine

The use of High Content Screening in Stem Cell Therapy and Research

Rod Bensen, Ph.D, Imagen Biotech

BIogRAphy:Robert Judson received his undergraduate degree in molecular biology and biochemistry from Wesleyan University in 2005. He spent two years as an IRTA fellow at the National Institute of Child Health and Human Development , NIH, in Bethesda, Maryland. There he researched the effect of nucleosome positioning on the targeting of HIV retrovirus and tf1 retrotransposon integration in the laboratory of Dr. Henry Levin.

In 2007 he began pursuing his PhD in the Biomedical Sciences program at UCSF. There he has been the recipient of the National Science Foundation Graduate Research Fellowship and the UC Cancer Research Coordinating Committee Fellowship. He is currently investigating the role of microRNAs in differentiation, de-differentiation and cell identity in the laboratory of Dr. Robert Blelloch.

BIogRAphy:Rod Bensen graduated from the University Sydney after an honours year majoring in neurobiological research. He then spent several years at Westmead hospital developing a Clinical database before moving to the UK where he completed his Ph.D. at the University of Manchester. Rod’s Ph.D. focussed on several cytological conditions that occur when cells commit to apoptosis.

In 2001, Rod obtained an 18 month Wellcome Showcase award for Innovative research where he explored the use of intrabodies tagged to fluorescent proteins to create an in-cyto immunoassay. The idea went by the acronym LADDERS (Live Antigen Detection Dual Epitope Reporter System). In 2003, Rod started working for AstraZeneca where he was employed in the Systems Biology group, Pathways. He left AZ in October 2007 to set up Imagen Biotech with his colleague Gareth Griffiths. Their aim was for Imagen Biotech to serve both the academic and industrial scientific community by providing a first class High Content Screening service.

There is a fundamental need for novel approaches to dissect important biological processes at a multi-gene systems level rather than one gene at a time. The unique features of microRNAs, being evolutionarily selected regulators of hundreds of mRNA targets with common physiological outcomes, places them in an optimal position to aid researchers in uncovering the proteins, pathways, and cellular functions mediating complex biological processes. Recent advances in our understanding of microRNA biology, as well as the development of tools to robustly over-express, inhibit, and determine the targets of microRNA s, are now allowing researchers to use microRNAs as tools to identify intricate signaling modules in cell biology. Using the de-differentiation of somatic cells into induced pluripotent stem cells as a case study, we will discuss how to set up and optimize a microRNA screen, verify hits, and follow these hits to information on development, pathways and mechanism.

The use of High Content Screening in Stem Cell Therapy and Research Automated fluorescence microscopy (High Content Analysis: HCA) is unique in that it is arguably the only method that offers high information content while also being high throughput. Therefore, it is an ideal tool for determining the culture conditions necessary to cause MSCs to undergo terminal differentiation because many treatment conditions can be rapidly analyzed. Similarly, the multiparametric readout of HCA means that several cell markers and cellular morphology can be determined from a single experiment. In our presentation we will demonstrate how HCA technology is the natural methodological choice for conducting stem cell related research.

In particular, this presentation will focus on the use of human Mesenchymal Stem Cells (MSC) in autologous cell therapies. These therapies involve the culture and expansion of MSCs for extended periods. Reports suggest extended MSC culture is

associated with spontaneous malignant cell transformation. MSC are also the likely cancer stem cell for bone tumors, notably osteosarcoma. We have examined whether non-transformed late passage MSC can be distinguished from transformed cells during culture expansion. We have also assessed MSCs taken from other patients and used HCA to quantify a panel of phenotypic markers which show variations in patient MSCs. Cell phenotypes established using these techniques will help clinical programmes develop release criteria for MSC therapies, especially to ensure their safe application.

Practical Applications of Stem cells for drug screening and toxicity testing.

Amy Sinor-Anderson, Ph.D., Thermo Fisher Scientific

Embryonic stem cells (ES) provide a valuable tool for both drug discovery and cell replacement therapies. One can use ES cells to generate large numbers of specific cell types needed to carry out screening campaigns for drug discovery and to understand basic biology of diseased states. This ability to generate enough relevant material is critical when studying certain diseases such as neurodegenerative disorders, where the traditional source material is typically primary tissue from animals. For instance, obtaining large numbers of ALS affected motor neurons for screening campaigns can be hindered by access to enough material. The differentiation of ES cells into motor neurons, however, has shown great promise as a viable method to alleviate this roadblock and provide access to the cultures required. Researchers are now able to take this one step further by generating disease specific cell types from human origin through reprogramming of somatic cells into induced pluripotent cells (iPS). Generating multiple iPS cell lines from patients can aid in profiling lead compounds in the drug discovery process. Currently, a major difficulty in the field is the availability of uniform methods and reagents for growing, expanding, differentiating, and characterizing such cells. In this talks we will discuss Thermo Scientific validated solutions that provide an optimized approach for the growth, expansion, differentiation, and characterization of ES and iPS cells with a high degree of confidence.

BIogRAphy:Amy Sinor is a Senior Research Scientist at Thermo Fisher Scientific, in the Biosciences division, focusing on developing products for stem cell research. Prior to joining, Thermo Fisher Scientific, she was a scientist at Curis Inc and the Harvard Stem Cell Institute. At Curis, she conducted small molecule screens in embryonic stem cell derived motor neurons to identify possible candidates to promote motor neuron survival using high content analysis. At Harvard Stem Cell Institute, she continued her work from Curis to identify small molecule candidates that promoted the up-regulation of a particular protein that was decreased during embryonic development.

Amy’s scientific interest has primarily focused on Neuroscience/Stem Cell research. Her initial research focused on studying the pharmacology of glutamate transporters in glial cultures and understanding the effects of ischemia/hypoxia on neuronal cultures and the mechanisms for protection. During her graduate work at the University of Pittsburgh, she studied the role of Akt, a serine-theronine kinase on the regulation of telencephalic progenitors/stem cells.

Amy earned a Ph.D. in Neurobiology from the University of Pittsburgh School of Medicine and received her B.A. in Biology and Chemistry from Skidmore College.

BIogRAphy:Paul Price has been active in the areas of cell culture and media development since 1958. He received his PhD from the George Washington University with majors in Cell Biology, Biochemistry and Virology and is currently a cell culture and media design consultant to Thermo Fisher Scientific. He was previously the Chief Scientific Officer of D-Finitive Cell Technologies (DCT), a specialty cell media company. Dr. Price has been on the Scientific Advisory Board and a consultant to several cell culture media and reagents companies and is an Adjunct Professor at the Medical University of South Carolina, Department of Regenerative Medicine. He was a Branch Chief in the Center for Infectious Diseases at the CDC and later founder and Executive Vice-President of Hycor Biomedical (later called Stratagene).

As part of R&D at GIBCO Invitrogen/Life Technologies, he helped design and develop much of their specialized media for the culture of neurons, stem cells and other mammalian cells, including those used in Bio-Production. As CSO of DCT, he developed numerous animal-origin-free and xeno-free formulations for many of the same cell types. Dr. Price has been an active member of the Society for In Vitro Biology since 1959 and the Society President from 2006-2008. He has been a reviewer for In Vitro and other journals and an outside expert for grant reviews for the NCI.

BIogRAphy:Dr. Cindy Neeley has been with Thermo Fisher Scientific for the past four years. She currently holds a position as Field Technical Specialist for cell culture product at the Laboratory Consumables Division. Prior to joining Thermo Fisher Scientific, Dr. Neeley received her Doctor’s degree in Immunology and Microbiology in 1996. She completed her post-doctoral training and held a position of Senior Research Scientist at University of Michigan. Dr. Neeley was recruited as R&D Manager for OptiCell product line by BioCrystal Ltd in 2003. The OptiCell business was later acquired by Thermo Fisher Scientific.

Applications of an Innovative Feeder-Free Plastic Surface for the Expansion of Human ES and IPS Cells

Cindy Neeley, Ph.D., Thermo Fisher Scientific

The concerns over contaminants from animal components and batch-to-batch variability of the coating matrices for human stem cells culture have significantly hindered the usage of this type of surface for translational and clinical applications. The Thermo Scientific Nunclon Vita is an energy-treated polystyrene surface free of animal components. It enables culture of human stem cells without matrix or feeder layers. Human embryonic stem (ES) cells are grown directly on the surface in conditioned media containing ROCK-inhibitor, and can be sustained for more than ten passages without signs of differentiation. During this presentation, we will also present data that demonstrate karyotypic normality, pluripotent status, and induced differentiation to pancreatic endoderm of human ES cells cultured on Nunclon Vita surface.

Media design for stem cell expansion, differentiation and preservation.

Paul Price, Ph.D., Thermo Fisher Scientific

The cell culture medium is a dynamic mixture consisting of amino acids, vitamins, a source of energy, growth factors, trace minerals and other components in a buffered salt solution. Each component has a shelf life, sensitivity to the physical environment and a variety of interactive and break-down products. The Classical mammalian cell culture formulations require further supplementation with a protein source such as serum and were designed using cancer-derived cell lines. Serum supplemented formulations for mammalian cells can be very sub-optimal for the growth of ES and other stem cells. This seminar will start with a very short history of media development and the role and problems associated with some of the key basic components of the medium and then show how we have been able to progress to formulations allowing for the growth of normal functional cell types in both 2-D and 3-D formats. Emphasis will be placed on how the cell culturist can reduce apoptosis by controlling the physical environment and the production of ammonia and free radicals through the optimization of the media formulation and the proper handling of the cells.