Optimized Approaches for Generation of Integration-free iPSCs from Human Urine-Derived Cells with Small Molecules and Autologous Feeder.
Optimized Approaches for Generation of Integration-free iPSCs from Human Urine-Derived Cells with Small Molecules and Autologous Feeder.
复制标题
从具有小分子和自体饲养层的人尿来源细胞生成无整合 iPSC 的优化方法
DOI：
10.1016/j.stemcr.2016.04.001
复制
发表时间：
2016-05-10
影响因子：
5.9
通讯作者：
Pan G
中科院分区：
文献类型：
--
作者：
Li D;Wang L;Hou J;Shen Q;Chen Q;Wang X;Du J;Cai X;Shan Y;Zhang T;Zhou T;Shi X;Li Y;Zhang H;Pan G
从人尿衍生细胞（hUC）产生诱导多能干细胞（iPSC）提供了一种方便且非侵入性的方式来获得患者特异性iPSC。然而，许多分离的hUC表现出非常差的增殖并且难以重编程。在这项研究中，我们优化了增殖非常差的hUC的重编程方法。我们在此报道了含有环匹非亭-a（一种P53抑制剂）、A-83-01、CHIR 99021、thiazovivin、NaB和PD 0325901的化合物混合物显著提高了hUC的重编程效率（170倍以上）。此外，我们表明，用自体hUC饲养细胞替代基质胶可以克服由于在重编程因子递送期间发生的大量细胞死亡而导致的重编程失败。总之，我们描述了改进的方法以使iPSC能够从否则难以重编程的hUC生成，这是储存患者特异性iPSC的宝贵资产。SM处理显著增强hUC的重编程用自体hUC替代Matrigel作为饲养层促进重编程细胞依赖性重编程策略的选择对于储存iPSC系是有用的在这篇文章中，Pan G，Zhang H，Li Y及其同事表明，在小分子的帮助下，增殖不良的hUC可以被重编程（A-83-01，Chir，Tzv，CPFT-a，NaB，PD）和自体UC饲养细胞。使用小分子混合物和自体饲养细胞的方法显着提高了重编程效率（170倍以上），并能够从具有不同增殖状态的hUC生成iPSC。
Generation of induced pluripotent stem cells (iPSCs) from human urine-derived cells (hUCs) provides a convenient and non-invasive way to obtain patient-specific iPSCs. However, many isolated hUCs exhibit very poor proliferation and are difficult to reprogram. In this study, we optimized reprogramming approaches for hUCs with very poor proliferation. We report here that a compound cocktail containing cyclic pifithrin-a (a P53 inhibitor), A-83-01, CHIR99021, thiazovivin, NaB, and PD0325901 significantly improves the reprogramming efficiency (170-fold more) for hUCs. In addition, we showed that replacement of Matrigel with autologous hUC feeders can overcome the reprogramming failure due to the massive cell death that occurs during delivery of reprogramming factors. In summary, we describe improved approaches to enable iPSC generation from hUCs that were otherwise difficult to reprogram, a valuable asset for banking patient-specific iPSCs. SM treatment significantly enhances the reprogramming of hUCs Replacement of Matrigel with autologous hUCs as feeder facilitates reprogramming Selection of cell-dependent reprogramming strategy is useful for banking iPSC lines In this article, Pan G, Zhang H, Li Y, and colleagues show that poorly proliferating hUCs could be reprogrammed with the aid of small molecules (A-83-01, Chir, Tzv, CPFT-a, NaB, PD) and autologous UC feeders. The approaches using a small-molecule cocktail and autologous feeder cells significantly improves the reprogramming efficiency (170-fold more) and enable iPSC generation from hUCs with different proliferation states.