Optimizing the method for generation of integration-free induced pluripotent stem cells from human peripheral blood

优化从人外周血中产生无整合诱导性多能干细胞的方法

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作者:Haihui Gu, Xia Huang, Jing Xu, Lili Song, Shuping Liu, Xiao-Bing Zhang, Weiping Yuan, Yanxin Li

Background

Generation of induced pluripotent stem cells (iPSCs) from human peripheral blood provides a convenient and low-invasive way to obtain patient-specific iPSCs. The episomal vector is one of the best approaches for reprogramming somatic cells to pluripotent status because of its simplicity and affordability. However, the efficiency of episomal vector reprogramming of adult peripheral blood cells is relatively low compared with cord blood and bone marrow cells.

Conclusions

With this optimized protocol, a valuable asset for banking patient-specific iPSCs has been established.

Methods

In the present study, integration-free human iPSCs derived from peripheral blood were established via episomal technology. We optimized mononuclear cell isolation and cultivation, episomal vector promoters, and a combination of transcriptional factors to improve reprogramming efficiency.

Results

Here, we improved the generation efficiency of integration-free iPSCs from human peripheral blood mononuclear cells by optimizing the method of isolating mononuclear cells from peripheral blood, by modifying the integration of culture medium, and by adjusting the duration of culture time and the combination of different episomal vectors. Conclusions: With this optimized protocol, a valuable asset for banking patient-specific iPSCs has been established.

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