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无饲养层条件下从红细胞中生成无转基因的人诱导多能干细胞。

Generation of transgene-free human induced pluripotent stem cells from erythroblasts in feeder-free conditions.

机构信息

Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland.

Laboratory of Neuroimmunology, Neuroscience Research Centre, Department of Clinical Neurosciences, Lausanne University Hospital (CHUV) and University of Lausanne, Lausanne, Switzerland.

出版信息

STAR Protoc. 2022 Aug 17;3(3):101620. doi: 10.1016/j.xpro.2022.101620. eCollection 2022 Sep 16.

Abstract

This protocol describes the generation and characterization of human induced pluripotent stem cells (hiPSCs) from erythroblasts. A key difference with classical protocols is the reprogramming of erythroblasts from a simple blood draw as opposed to fibroblasts/keratinocytes, which requires a biopsy. Moreover, working with erythroblasts ensures that no recombination of the TCR/BCR genes occurs, as opposed to T cells and whole peripheral blood mononuclear cells-based approaches. Last, this approach uses non-integrative episomes ensuring no integration of transgenes into the hiPSCs genome. For complete details on the use and execution of this protocol, please refer to Perriot et al. (2018).

摘要

本方案描述了从红细胞生成人诱导多能干细胞(hiPSC)的方法及其鉴定。与经典方案相比,其主要区别在于本方案中通过简单的血液采集而非活检来获得成纤维细胞/角质细胞进行重编程。此外,使用红细胞可以确保 TCR/BCR 基因不发生重组,这与 T 细胞和基于整个外周血单核细胞的方法不同。最后,这种方法使用非整合的附加体,确保转基因不会整合到 hiPSC 的基因组中。有关该方案使用和实施的完整详细信息,请参见 Perriot 等人(2018 年)。

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