Human embryonic stem cell (hESC) research depends upon the ability of these cells to remain pluripotentand genetically stable in culture as a self-renewing population. This stability and functionality must also bemaintained after directed differentiation into multiple cellular lineages. Current culture conditions for hESCsare sub-optimal and this can lead to acquisition of chromosome abnormalities, sporadic differentiation,genomic alterations and decreased cell viability. Before hESCs will be useful clinically, standard protocolsmust be developed to ensure accurate and repeatable methods for hESC maintenance, culture anddifferentiation. Although most of the NIH approved lines share common characteristics, a wide range ofculture conditions have been published, which makes comparisons of different methods difficult.The hESC Core Facility (Core B) is developing standardized hESC culture methods with the goal of assistingresearchers in this program project with hESC culture techniques. The core will provide expertise ingrowing, splitting, monitoring and determining valid protocols for hESC culture methods. In addition, the corewill develop crucial reagents for each of the projects, as detailed in Projects 1-3. Core B will provide a meansfor project integration and comparison, and also train investigators as new technology develops. Core B willprovide novel biosynthetic culture surfaces to optimize hESC growth in both undifferentiated anddifferentiated states. Thus, Core B will provide both service and experimental activities for the benefit of theprogram project.
The Specific Aims of the Stem Cell Core are:
Specific Aim 1. To culture, bank and distribute hESCs for the program projectSpecific Aim 2. Monitor differences over prolonged culture conditions with multiple hESC cell linesSpecific Aim 3. Optimize growth conditions with multiple hESC lines as needed for each projectSpecific Aim 4. Provide hESCs with stable vector integration and lysates as needed per projectSpecific Aim 5. Develop new directions in synthetic materials and architecturesSpecific Aim 6. Integrate projects and facilitate transfer of information between each project
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