Streamlined Banking of Human Pluripotent Stem Cells Through Molecular Authenticat

DESCRIPTION (provided by applicant): The recent creation of human induced pluripotent stem cells (iPSC) from reprogrammed or de-differentiated adult cells has changed the approach and attitude of many to pluripotent stem cell research. These new iPSC are potentially a valuable source of in vitro models of complex, polygenic human diseases. They may find an important role in the discovery of new drugs and therapies. They are also a potential downstream source of cells for autologous cell therapy. However, these new cells are still subject to some of the same roadblocks that have been standing in the way of advancing human embryonic stem cell (hESC) research. These cells are inherently difficult to maintain in culture, they require meticulous quality control and standardization, and they must be made readily available to the research community. Cell culture has played a central and critical role in the advancement of both basic and translational research. As a result, scientists and institutions have come to recognize the advantages of maintaining cell banks in order to support the development and maintenance of healthy cell lines. In this application, we propose to create a streamlined human pluripotent stem cell (hPSC - hESC and iPSC) banking system in which we will optimize the authentication and quality control (QC) processes of hPSC lines. Human iPSC and human ESC share many common properties - including culture requirements, marker expression, and pluripotency. We propose to use three human iPSC lines as proof-of-principle for hPSC banking. These are potentially interesting and important cell lines, especially for the hematopoietic and mesenchymal stem cell fields. This system will be cost effective and will strive for self-sufficiency. We will greatly enhance the turnaround time for making the banks of cells available by employing a characterization scheme mainly comprised of molecular assays. The proposed assays are less expensive (requiring less cellular material) and quicker (having shorter run times) than traditional assays. We will validate the new assays by comparing the results with those obtained by traditional tests being used now - including immunophenotyping and G-band karyotyping. Once accomplished, we will have taken a large step forward toward realizing the full potential of these stem cells. This is a forward-looking application with a vision to the near future need for banking hundreds of hPSC lines. PUBLIC HEALTH RELEVANCE: In this proposal we are laying the groundwork for an hPSC bank that will be able to respond quickly to the impending flood of hESC and iPSC lines. By validating a set of molecular authentication assays we will have taken the first step toward an efficient banking system. The ultimate endpoint is an hPSC bank that can quickly and economically accession and make important cell lines available to the research community.