Chemical ablation of tumor-initiating human pluripotent stem cells
Chemical ablation of tumor-initiating human pluripotent stem cells
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DOI：
10.1038/nprot.2014.050
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发表时间：
2014-03-01
期刊：
影响因子：
14.8
通讯作者：
Benvenisty, Nissim
中科院分区：
文献类型：
--
作者：
Ben-David, Uri;Benvenisty, Nissim
人多能干细胞（hPSC）的致瘤性被广泛认为是阻碍其在再生医学中应用的主要障碍。该方案描述了从单层培养物中化学消除肿瘤起始hPSC的两种有效且稳健的方法。该方案详细说明了如何维持和分化hPSC，如何将化学抑制剂应用于hPSC及其分化后代的培养物，以及如何使用体外和体内测定来评估所得细胞培养物的纯度。它还描述了如何挽救细胞毒性效应。消除和补救试验可在3-5天内完成，体外评估需要另一天，体内评估需要额外长达12周。
The tumorigenicity of human pluripotent stem cells (hPSCs) is widely acknowledged as a major obstacle that withholds their application in regenerative medicine. This protocol describes two efficient and robust ways to chemically eliminate the tumorinitiating hPSCs from monolayer culture. The protocol details how to maintain and differentiate hPSCs, how to apply chemical inhibitors to cultures of hPSCs and their differentiated progeny, and how to assess the purity of the resultant cell cultures using in vitro and in vivo assays. It also describes how to rescue the cytotoxic effect. The elimination and the rescue assay can be completed within 3-5 d, the in vitro assessment requires another day, and the in vivo assessment requires up to 12 additional weeks.