Enhanced plating efficiency of trypsin-adapted human embryonic stem cells is reversible and independent of trisomy 12/17
Enhanced plating efficiency of trypsin-adapted human embryonic stem cells is reversible and independent of trisomy 12/17
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DOI：
10.1089/clo.2007.0064
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发表时间：
2008-03-01
期刊：
影响因子：
--
通讯作者：
Daley, George Q.
中科院分区：
文献类型：
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作者：
Chan, Elayne M.;Yates, Frank;Daley, George Q.
人胚胎干细胞（human embryonic stem cells，hESC）可以大量培养，但存在染色体畸变。为了保持其遗传完整性，hESC通常在传代期间保持为细胞聚集体或团块。然而，团块传代阻碍了大规模培养并使单细胞克隆的分离复杂化。为了促进hESC的遗传修饰克隆的分离，同时保持其遗传完整性，我们采用胰蛋白酶单细胞传代短暂的时间，然后返回到丛传代进行长期维护。我们观察到，适应胰蛋白酶作为单细胞传代是一个适应性过程，其中超过三至四次传代显著提高了平板接种效率。然而，胰蛋白酶通道与染色体12和17的异常。然而，胰蛋白酶传代的hESC的高平板接种效率是可逆的表型，无论染色体异常，表明表观遗传事件是表型转换的原因。
Human embryonic stem cells (hESCs) can be cultured abundantly and indefinitely, but are subject to accumulations of chromosomal aberrations. To preserve their genetic integrity, hESCs are commonly maintained as cell aggregates or clumps during passaging. However, clump passaging hinders large-scale culture and complicates the isolation of single cell clones. To facilitate the isolation of genetically modified clones of hESCs while preserving their genetic integrity, we employed trypsin single-cell passaging for brief periods before returning to clump passaging for long-term maintenance. We observed that accommodation to trypsin passage as single cells is an adaptive process where over three to four passages considerably increases the plating efficiency. However, trypsin passage was associated with abnormalities of chromosomes 12 and 17. Nevertheless, the high plating efficiency of trypsin passaged hESCs is a reversible phenotype, regardless of chromosomal abnormalities, suggesting that epigenetic events are responsible for the switch in phenotype.