Wu Yuehong, Shu Jianhong, He Chengwen, Li Min, Wang Yujiong, Ou Wenbin, He Yulong
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China; Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, College of Life Science, Ningxia University, Yinchuan, Ningxia, China.
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China.
Theriogenology. 2016 Jan 15;85(2):302-14. doi: 10.1016/j.theriogenology.2015.09.020. Epub 2015 Sep 25.
Previous studies reported that Rho-associated kinase inhibitor Y27632 markedly diminishes human embryonic stem cell and induced pluripotent stem cell (iPSC) dissociation-induced apoptosis and increases cloning efficiency in a feeder-free culture system. However, the mechanisms by which Y27632 protects pluripotent stem cells from apoptosis remain unknown. In the present study, we tested the effects of Y27632 on single dissociated marmoset iPSCs in a feeder-free culture. The results showed that Y27632 promoted the number of cells proliferating after passage by single-cell dissociation in a dose-dependent manner. The Rho-associated kinase inhibitor Y27632 markedly increased the cloning efficiency of marmoset iPSCs without affecting their karyotype and the expression of pluripotency markers. Meanwhile, Y27632 markedly diminished apoptosis of the marmoset iPSCs under even more severe conditions by suppressing the expression and activity of caspase 3. Taken together, the present results suggest that this reagent is effective in improving the cultural system of primate iPSCs.
先前的研究报道，Rho相关激酶抑制剂Y27632可显著减少人胚胎干细胞和诱导多能干细胞（iPSC）解离诱导的凋亡，并在无饲养层培养系统中提高克隆效率。然而，Y27632保护多能干细胞免于凋亡的机制仍不清楚。在本研究中，我们测试了Y27632对无饲养层培养中单个解离的狨猴iPSC的影响。结果表明，Y27632以剂量依赖的方式促进了单细胞解离传代后增殖的细胞数量。Rho相关激酶抑制剂Y27632显著提高了狨猴iPSC的克隆效率，而不影响其核型和多能性标志物的表达。同时，Y27632通过抑制caspase 3的表达和活性，在更严峻的条件下显著减少了狨猴iPSC的凋亡。综上所述，目前的结果表明该试剂在改善灵长类iPSC培养系统方面是有效的。
