西亚试剂：iPS细胞潜力的细胞标记

 以前试图分离正处在成功的“诱导多能干”(iPS)细胞重新编程途中之细胞的努力一直基于这样一个假设：细胞渐进性地失去供体细胞身份并逐渐获得iPS细胞特性。在这项研究中，Marius Wernig及同事识别出了在该过程早期出现的表面标记，它们在最初的成纤维细胞或最终产生的多能细胞中都没有表达。通过分离和分析具有这些标记的细胞(包括CD73、CD49d和CD200)的表达，作者识别出了重新编程所需的新的转录调控因子，从而获得了对这一过程机制的认识。

原文标题：Early reprogramming regulators identified by prospective isolation and mass cytometry

原文摘要：In the context of most induced pluripotent stem (iPS) cell reprogramming methods, heterogeneous populations of non-productive and staggered productive intermediates arise at different reprogramming time points1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11. Despite recent reports claiming substantially increased reprogramming efficiencies using genetically modified donor cells12, 13, prospectively isolating distinct reprogramming intermediates remains an important goal to decipher reprogramming mechanisms. Previous attempts to identify surface markers of intermediate cell populations were based on the assumption that, during reprogramming, cells progressively lose donor cell identity and gradually acquire iPS cell properties1, 2, 7, 8, 10. Here we report that iPS cell and epithelial markers, such as SSEA1 and EpCAM, respectively, are not predictive of reprogramming during early phases. Instead, in a systematic functional surface marker screen, we find that early reprogramming-prone cells express a unique set of surface markers, including CD73, CD49d and CD200, that are absent in both fibroblasts and iPS cells. Single-cell mass cytometry and prospective isolation show that these distinct intermediates are transient and bridge the gap between donor cell silencing and pluripotency marker acquisition during the early, presumably stochastic, reprogramming phase2. expression profiling reveals early upregulation of the transcriptional regulators Nr0b1 and Etv5 in this reprogramming state, preceding activation of key pluripotency regulators such as Rex1 (also known as Zfp42), Dppa2, Nanog and Sox2. Both factors are required for the generation of the early intermediate state and fully reprogrammed iPS cells, and thus represent some of the earliest known regulators of iPS cell induction. Our study deconvolutes the first steps in a hierarchical series of events that lead to pluripotency acquisition.

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