Center for Regenerative Medicine in Barcelona, Barcelona, Spain.
Nat Protoc. 2010 Apr;5(4):647-60. doi: 10.1038/nprot.2010.9. Epub 2010 Mar 11.
The generation of patient-specific induced pluripotent stem cells (iPSCs) offers unprecedented opportunities for modeling and treating human disease. In combination with gene therapy, the iPSC technology can be used to generate disease-free progenitor cells of potential interest for autologous cell therapy. We explain a protocol for the reproducible generation of genetically corrected iPSCs starting from the skin biopsies of Fanconi anemia patients using retroviral transduction with OCT4, SOX2 and KLF4. Before reprogramming, the fibroblasts and/or keratinocytes of the patients are genetically corrected with lentiviruses expressing FANCA. The same approach may be used for other diseases susceptible to gene therapy correction. Genetically corrected, characterized lines of patient-specific iPSCs can be obtained in 4-5 months.
患者特异性诱导多能干细胞（iPSC）的产生为人类疾病的建模和治疗提供了前所未有的机会。与基因治疗相结合，iPSC 技术可用于生成无疾病的祖细胞，这些祖细胞可能对自体细胞治疗具有潜在的意义。我们解释了一种使用 OCT4、SOX2 和 KLF4 的逆转录病毒转导，从范可尼贫血患者的皮肤活检中可重复产生遗传纠正 iPSC 的方案。在重编程之前，使用表达 FANCA 的慢病毒对患者的成纤维细胞和/或角质形成细胞进行基因纠正。该方法也可用于其他易受基因治疗纠正的疾病。可以在 4-5 个月内获得遗传纠正的、具有特征的患者特异性 iPSC 系。
