State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Center for Stem Cell Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, 200093, China.
Department of Transfusion Medicine, Shanghai Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai, 200433, China.
Stem Cell Res Ther. 2018 Jun 15;9(1):163. doi: 10.1186/s13287-018-0908-z.
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.
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.
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.
With this optimized protocol, a valuable asset for banking patient-specific iPSCs has been established.
从人外周血生成诱导多能干细胞（iPSCs）提供了一种方便、低侵袭性的方法来获得患者特异性 iPSCs。由于其简单性和可负担性，附加体载体是将体细胞重编程为多能状态的最佳方法之一。然而，与脐带血和骨髓细胞相比，附加体载体重编程成人外周血细胞的效率相对较低。
在本研究中，通过附加体技术建立了无整合的人 iPSCs。我们优化了单核细胞分离和培养、附加体载体启动子以及转录因子的组合，以提高重编程效率。
在这里，我们通过优化从外周血中分离单核细胞的方法、修改培养基的整合、调整培养时间的持续时间以及不同附加体载体的组合，提高了无整合 iPSCs 从人外周血单个核细胞中的生成效率。
通过这个优化方案，建立了一个有价值的用于储存患者特异性 iPSCs 的方法。
