从外周血单核细胞生成患者特异性 iPSC 系的方案。
A protocol for the generation of patient-specific iPSC lines from peripheral blood mononuclear cells.
机构信息
The Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang 310058, China; Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China.
The Children's Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, Zhejiang 310058, China.
出版信息
STAR Protoc. 2022 Sep 16;3(3):101530. doi: 10.1016/j.xpro.2022.101530. Epub 2022 Jul 13.
The current procedure thoroughly explains how to reprogram induced pluripotent stem cells (iPSCs) from the patient's peripheral blood mononuclear cells (PBMCs), a less invasive source; e.g., somatic cells. We describe how to isolate PBMCs and reprogram them into iPSCs by electroporation. Furthermore, we provide an alternative approach to generating iPSC using Geltrex or Matrigel matrix to replace MEF-feeder. The challenge of this process is the relatively lower cell survival rates of PBMCs due to the damage of electroporation. For complete details on the use and execution of this protocol, please refer to Hu et al. (2021).
目前的程序详细说明了如何从患者外周血单核细胞（PBMCs）中重新编程诱导多能干细胞（iPSCs），这是一种侵袭性更小的来源，例如体细胞。我们描述了如何分离 PBMCs 并通过电穿孔将其重新编程为 iPSCs。此外，我们提供了一种使用 Geltrex 或 Matrigel 基质代替 MEF 饲养层生成 iPSC 的替代方法。该过程的挑战是由于电穿孔的损伤，PBMCs 的细胞存活率相对较低。有关该方案使用和执行的详细信息，请参阅 Hu 等人（2021 年）。
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