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目的探讨碱性成纤维细胞因子(basic fibroblast growth factor,bFGF)浓度对人类胚胎干细胞(human embry-onicstem cells,hESCs)特性的影响。方法建立化学限定性hESCs培养体系,以有饲养细胞培养体系作为参照,比较4、40ng/ml和100ng/mlbFGF对hESCs多能性标记、细胞周期、细胞凋亡和DNA完整性的影响。结果①低浓度bFGF(4ng/ml)足以维持hESCs在化学限定性培养基(基础培养基DMEM/F12,N2,B27,简称NB体系)中培养;②bFGF可以抑制hESCs细胞凋亡,且具有剂量依赖性,而对hESCs细胞周期没有影响;③bFGF有助于hESCsDNA完整性维持,在NB体系加入4ng/mlbFGF时,hESCs在DNA水平上存在75个异常区域,而在加入40ng/mlbFGF时,只存在1个异常区域。结论高剂量bFGF有助于hESCs体外快速扩增和hESCsDNA完整性维持。提高bFGF浓度为hESCs体外培养所必需。
Objective To investigate the effect of basic fibroblast growth factor (bFGF) on the characteristics of human embryonic stem cells (hESCs). Methods The chemosensory hESCs culture system was established. The effects of 4,40ng / ml and 100ng / ml bFGF on the pluripotency markers, cell cycle, apoptosis and DNA integrity of hESCs were compared with the culture system of feeder cells. Results ① Low concentration of bFGF (4ng / ml) was enough to maintain hESCs in chemically defined medium (basal medium DMEM / F12, N2, B27, NB system for short); bFGF could inhibit hESCs apoptosis in a dose- While hESCs had no effect on the cell cycle. BFGF contributed to the maintenance of the integrity of hESCs DNA. In the NB system with 4ng / ml bFGF, there were 75 abnormal regions of DNA at the level of hESCs, but only 1 Abnormal area. Conclusion High-dose bFGF contributes to the rapid expansion of hESCs in vitro and the maintenance of hESCs DNA integrity. Increasing bFGF concentration is necessary for in vitro culture of hESCs.