(251d) Combinatorial Development of Synthetic Polymeric Substrates for Clonal Growth of Human Pluripotent Stem Cells

Human embryonic stem (ES) and induced pluripotent stem (iPS) cells survive and self-renew poorly when dissociated into single cells, making the isolation of genetically modified hES and hiPS cells a cumbersome process. We hypothesized that substrate-mediated signaling influences clonal growth and could be engineered to enhance this process. To this end, a library of cell-compatible protein-coated polymer substrates were utilized in a miniaturized, high-throughput nanoliter format. These nanoliter polymer substrate arrays were then screened for colony growth from a single hES cell. After screening thousands of protein-coated polymers, multiple ?hits' indicated that a specific range of polymeric structures could promote single cell growth. Propagation of hES cells on these substrates showed that the resulting colonies retained a pluripotent phenotype, even after subsequent passaging. These hits also were utilized with hiPS cells. By identifying substrates which support clonal growth and self-renewal of hES and hiPS cells, this work can help establish robust, broadly useful culture platforms to accelerate progress in manipulating human pluripotent cells in vitro, which will allow for the systematic analysis of mechanisms involved in human development and disease.