This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.To develop a media formulation and protocol for the human ES cell culture that will improve human ES cell viability and promote uniformity, consistency and ease of use across a variety of users in different locations. Some of the specific objectives necessary to accomplish this goal are to: 1) optimize the physiochemical environment, 2) optimize the basal media formulation and 3) eliminate undefined media components. In this budget period, we were able to develop a both a short-term and intermediate screening method that will significantly reduce the cost and time required for future optimization studies. Using this new assay system we have already identified several factors that contribute to self-renewal of human ES cells in a defined culture systems. This data will serve as the basis for future grant applications. Additionally the physiochemical environment for human ES cells, including pH, osmolarity and gas amosphere, was optimized. Lastly more than 90 individual factors were screened for their effect on human ES cell competence (including cell morphology, proliferation and spontaneous differentiation rates) This research used WNPRC resources and federally approved hES cell lines.
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