Non-Xenogeneic Expansion And Definitive Endoderm Differentiation Of Human Pluripotent Stem Cells In An Automated Bioreactor
BIOTECHNOLOGY AND BIOENGINEERING(2021)
摘要
Scalable processes are requisite for the robust biomanufacturing of human pluripotent stem cell (hPSC)-derived therapeutics. Toward this end, we demonstrate the xeno-free expansion and directed differentiation of human embryonic and induced pluripotent stem cells to definitive endoderm (DE) in a controlled stirred suspension bioreactor (SSB). Based on previous work on converting hPSCs to insulin-producing progeny, differentiation of two hPSC lines was optimized in planar cultures yielding up to 87% FOXA2(+)/SOX17(+) cells. Next, hPSCs were propagated in an SSB with controlled pH and dissolved oxygen. Cultures displayed a 10- to 12-fold increase in cell number over 5-6 days with the maintenance of pluripotency (>85% OCT4(+)) and viability (>85%). For differentiation, SSB cultures yielded up to 89% FOXA2(+)/SOX17(+) cells or similar to 8 DE cells per seeded hPSC. Specification to DE cell fate was consistently more efficient in the bioreactor compared to planar cultures. Hence, a tunable strategy is established that is suitable for the xeno-free manufacturing of DE cells from different hPSC lines in scalable SSBs. This study advances bioprocess development for producing a wide gamut of human DE cell-derived therapeutics.
更多查看译文
关键词
definitive endoderm (DE) differentiation, human pluripotent stem cells (hPSCs), pluripotent expansion, stirred suspension bioreactor (SSB), xeno-free
AI 理解论文
溯源树
样例
生成溯源树,研究论文发展脉络
Chat Paper
正在生成论文摘要