Successful therapeutic application of human pluripotent stem cells (hPSCs) requires their large-scale generation as stable pluripotent cells and their differentiated functional progeny. We adopted the stirred-suspension bioreactor platform for propagating hPSCs in a chemically defined environment under self-renewing and directed differentiation modes. For microcarrier culture, the low adhesion of hPSCs on the beads is a significant bottleneck. Optimal conditions were identified and compared to our previous methodology, the efficiency of microcarrier seeding increased from 30% to over 50%. The distribution of cells on microcarriers was more even resulting in aggregate-free cultures. The concentration of cultured hPSCs under xeno-free conditions increased 30-40 fold over 8 days and hPSC pluripotency was ascertained by qPCR and flow cytometry. When hPSCs were subjected to differentiation they displayed markers characteristic of different lineages. Our studies support the use of stirred-suspension microcarrier bioreactors for the expansion of hPSCs under chemically defined conditions.

Propagation and directed differentiation of human pluripotent stem cells in stirred-suspension culture

243rd National Meeting (2012)

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