科研之友
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摘要
Purpose Development of improved protocols for differentiating induced pluripotent stem (iPS) cells into hepatic cells is an important step toward their use in the field of hepatology. Specifically, the number of different cytokines should be reduced to limit undesired effects and to reduce the cost of the process. In this report, we describe a simple method for directing human iPS cells to differentiate into hepatic cells using only two cytokines and a short incubation time. Methods A two-step protocol for differentiating iPS cells into hepatic cells was developed. A high dose of activin A was applied for 3 days to induce definitive endoderm formation. Subsequently, cells were treated with hepatocyte growth factor (HGF) for 5 days to generate hepatic cells. Differentiation was confirmed by immunostaining for differentiation markers. Albumin mRNA levels in differentiated hepatic cells generated using a previously tested three-step protocol that uses activin A, fibroblast growth factor (FGF)/bone morphogenetic protein (BMP), and HGF, and our new protocol were compared to determine the efficiency of differentiation. Results Our two-step protocol induced the differentiation of iPS cells into hepatic cells and required a shorter differentiation period than the previous three-step protocol. The differentiation efficiencies of the two protocols were comparable and the induced hepatic cells were functional. Conclusions Developing efficient induction and culture methods to generate more highly matured hepatocytes is essential for regenerative cell-based therapies. Our protocol provides a simple, cost-effective, and time-saving approach for generating hepatic cells from iPS cells.
- 出版日期2011-12
- 单位河北医科大学
