Role of caspase-3 inhibitor in induced anoikis of mesenchymal stem cells in vitro
Role of caspase-3 inhibitor in induced anoikis of mesenchymal stem cells in vitro
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DOI：
10.1007/s11596-007-0220-0
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发表时间：
2007-04-01
影响因子：
--
通讯作者：
Tang Xin
中科院分区：
文献类型：
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作者：
Feng Jianjun;Yang Shuhua;Tang Xin
本研究通过阻止骨髓间充质干细胞（MSCs）粘附于琼脂糖表面，建立MSCs悬浮液体外培养模型，探讨MSCs的失巢凋亡及caspase-3在失巢凋亡中的作用。将培养的MSCs随机分为3组：失巢凋亡组、caspase-3抑制剂组和对照组。实验前用1.5%琼脂糖包被培养皿;失巢凋亡组将MSCs放入预包被培养皿中;抑制剂组将caspase-3抑制剂和MSCs同时放入预包被培养皿中;对照组不进行干预。分别于2 h、6 h、12 h和24 h收集骨髓间充质干细胞。caspase-3荧光法和western blot法检测caspase-3活性的变化。流式细胞仪检测细胞凋亡率。失巢凋亡组和抑制剂组MSCs呈圆形，悬浮充分。Caspase-3荧光检测结果显示失巢凋亡组与对照组比较差异有统计学意义（P
By preventing mesenchymal stein cells (MSCs) from adhering to precoated agarose to create a model of MSC suspension in vitro, we investigated anoikis in MSCs and the role of caspase-3 in the anoikis. The cultured MSCs were randomly divided into 3 groups: the anoikis group, caspase-3 inhibitor group and control group. Before experiment, we coated dishes with 1.5 % agarose; in the anoikis group, MSCs were put into the precoated dishes; and in the inhibitor group, caspase-3 inhibitor and MSCs were also put into the precoated dishes; but there were not intervention in the control group. MSCs were collected at 2 h, 6 h, 12 h and 24 h. The alteration of caspase-3 activity was evaluated by caspase-3 fluorometric assay and western blot analysis. The apoptosis rates were detected by flow cytometry. MSCs were round and suspended sufficiently in the anoikis and inhibitor groups. Caspase-3 fluorometric assay showed that there were significant differences in statistics between the anoikis group and the others (P