Morgridge Institute for Research, Madison, Wisconsin, USA.
Nat Methods. 2011 May;8(5):424-9. doi: 10.1038/nmeth.1593. Epub 2011 Apr 10.
We re-examine the individual components for human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) culture and formulate a cell culture system in which all protein reagents for liquid media, attachment surfaces and splitting are chemically defined. A major improvement is the lack of a serum albumin component, as variations in either animal- or human-sourced albumin batches have previously plagued human ESC and iPSC culture with inconsistencies. Using this new medium (E8) and vitronectin-coated surfaces, we demonstrate improved derivation efficiencies of vector-free human iPSCs with an episomal approach. This simplified E8 medium should facilitate both the research use and clinical applications of human ESCs and iPSCs and their derivatives, and should be applicable to other reprogramming methods.
我们重新检查了人类胚胎干细胞（ESC）和诱导多能干细胞（iPSC）培养的各个组成部分，并制定了一个细胞培养系统，其中所有液体培养基、附着表面和分裂的蛋白质试剂都是化学定义的。一个主要的改进是缺乏白蛋白成分，因为以前动物源或人源白蛋白批次的变化一直困扰着人类 ESC 和 iPSC 培养，导致不一致。使用这种新的培养基（E8）和纤连蛋白涂层表面，我们展示了使用无载体的外源性方法提高了人类 iPSC 的衍生效率。这种简化的 E8 培养基应该有助于人类 ESC 和 iPSC 及其衍生物的研究和临床应用，并且应该适用于其他重编程方法。
