Division of Neuroscience, Stem Cells and Neurogenesis Unit, San Raffaele Scientific Institute, Milan, Italy.
Stem Cells. 2011 Nov;29(11):1684-95. doi: 10.1002/stem.726.
Induced pluripotent stem cell (iPSC) technology has provided researchers with a unique tool to derive disease-specific stem cells for the study and possible treatment of degenerative disorders with autologous cells. The low efficiency and heterogeneous nature of reprogramming is a major impediment to the generation of personalized iPSC lines. Here, we report the generation of a lentiviral system based on a microRNA-regulated transgene that enables for the efficient selection of mouse and human pluripotent cells. This system relies on the differential expression pattern of the mature form of microRNA let7a in pluripotent versus committed or differentiated cells. We generated microRNA responsive green fluorescent protein and Neo reporters for specific labeling and active selection of the pluripotent cells in any culture condition. We used this system to establish Rett syndrome and Parkinson's disease human iPSCs. The presented selection procedure represents a straightforward and powerful tool for facilitating the derivation of patient-specific iPSCs.
诱导多能干细胞（iPSC）技术为研究人员提供了一种独特的工具，可以从自体细胞中获得特定疾病的干细胞，用于退行性疾病的研究和可能的治疗。重编程的低效率和异质性是生成个性化 iPSC 系的主要障碍。在这里，我们报告了一种基于 microRNA 调控转基因的慢病毒系统的产生，该系统能够有效地选择小鼠和人类多能细胞。该系统依赖于 microRNA let7a 的成熟形式在多能性与定向分化或分化细胞中的差异表达模式。我们生成了 microRNA 反应性绿色荧光蛋白和 Neo 报告基因，用于在任何培养条件下对多能细胞进行特异性标记和主动选择。我们使用该系统建立了雷特综合征和帕金森病的人类 iPSC。所提出的选择程序代表了一种简单而强大的工具，可促进患者特异性 iPSC 的衍生。
