Eppley Institute for Research in Cancer & Allied Diseases & Department of Biochemistry & Molecular Biology, University of Nebraska Medical Center, Omaha, Nebraska 68198-6805, USA.
Regen Med. 2010 Sep;5(5):799-807. doi: 10.2217/rme.10.45.
Until recently, culturing human pluripotent stem cells was hampered by three prominent technical problems: a high degree of unwanted cellular stress when the cells are passaged, unacceptably low cloning efficiency and poor recovery of cryopreserved stocks. This review discusses recent developments that address these problems. A major focus of the review is the use of p160 Rho-associated coiled-coil kinase inhibitors for improving both the cloning efficiency and the recovery of cryopreserved human embryonic stem cells and human induced pluripotent stem cells. An underlying theme of this review is that the three problems have a common cause: separation of human pluripotent stem cells from one another increases cellular stress, which greatly decreases their viability unless special steps are taken.
直到最近,培养人类多能干细胞还受到三个突出的技术问题的阻碍:当细胞传代时,会产生高度不必要的细胞应激,克隆效率不可接受地低,以及冷冻保存品的复苏效果不佳。本文讨论了针对这些问题的最新进展。本文的一个主要重点是使用 p160 Rho 相关卷曲螺旋激酶抑制剂来提高克隆效率和冷冻保存的人类胚胎干细胞和人类诱导多能干细胞的复苏效果。本文的一个基本主题是,这三个问题有一个共同的原因:人类多能干细胞彼此分离会增加细胞应激,除非采取特殊措施,否则会大大降低其活力。