大鼠骨髓间充质干细胞的分离培养及rhEGF对其增殖的影响

Isolation and culture mesenchymal stem cells (MSCs) of rat and the effects of recombinant human epidermal growth factor (rhEGF) on MSCs proliferation in rats

目的分离培养大鼠骨髓间充质干细胞(MSCs),并探讨人重组表皮生长因子(rhEGF)对其体外增殖的影响。方法用贴壁培养法分离纯化大鼠骨髓MSCs,传代扩增,将生长良好的传代细胞分为rhEGF组和空白对照组,rhEGF组加20 ngrhEGF和含20%新生牛血清(FBS)的DMEM培养基,对照组只加含20%FBS的DMEM培养基,连续观察细胞形态及生长特性,绘制生长曲线。结果 2、4、6代细胞生长曲线基本相同,细胞倍增时间约34 h,方差分析P>0.05,三代细胞间无统计学差异。rhEGF组与空白对照组的数值,分别做重复测量方差分析,P<0.01,差异有统计学意义。结论骨髓MSCs体外稳定扩增,有很强的增殖能力,且rhEGF能促进其增殖。

Objective To isolate and culture mesenchymal stem cells（MSCs） in adult rat and study the effects of rhEGF on MSCs proliferation.Methods MSCs were isolated and purified by density gradient centrifugation.After successive subculture and amplification,the cells were separated into two groups,one group 20ng rhEGF ＋20% FBS/DMEM were added,another group only 20% FBS/DMEM were added.Morphology and proliferation characteristics were observed and the growth curves were described.Results The growth curves of passage 2,4,6 were quite similar,population doubling time was about 34 hours（P〉0.05）,there was no statistic difference among the three passages,the difference was statistically significant between the EGF group and the control group（P〈0.01）.Conclusion Marrow mesenchymal cells of adult rats are successfully isolated and they show stable expansion and active proliferation capacity in vitro.rhEGF can promote MSCs proliferation.