Epigenetics Programme, Babraham Institute, Cambridge, UK.
Methods Mol Biol. 2022;2416:29-37. doi: 10.1007/978-1-0716-1908-7_3.
Human pluripotent stem cells exist in naïve and primed states that recapitulate the distinct molecular and cellular properties of pre- and post-implantation epiblast cells, respectively. Naïve pluripotent stem cells can be captured directly from blastocysts but, more commonly, the cells are reprogrammed from primed cells in a process called "resetting". Several methods to achieve resetting have been described. Chemical resetting of primed cells to a naïve pluripotent state is one such method and has come to the forefront as a simple, efficient, and transgene-free method to induce naïve pluripotency. The process involves the transient application of a histone deacetylase inhibitor to initiate resetting, followed by the emergence of nascent naïve pluripotent stem cells in supportive conditions, and finally the stabilization and expansion of naïve pluripotent stem cell cultures. Here, a detailed protocol is provided for chemical resetting starting from plating primed cells until a stable culture of naïve pluripotent stem cells is established.
人类多能干细胞存在于原始态和起始态，分别再现了着床前和着床后上胚层细胞的独特分子和细胞特性。原始态多能干细胞可以直接从囊胚中捕获，但更常见的是，通过一种称为“重置”的过程从起始态细胞重新编程。已经描述了几种实现重置的方法。将起始态细胞化学重置为原始态多能性是这样一种方法，它作为一种简单、高效且无转基因的诱导原始多能性的方法而备受关注。该过程涉及短暂应用组蛋白去乙酰化酶抑制剂来启动重置，随后在支持条件下出现新生原始态多能干细胞，最后稳定和扩增原始态多能干细胞培养物。在这里，提供了一个从 plating 起始态细胞开始到建立稳定的原始态多能干细胞培养物的详细化学重置方案。
