碱性成纤维细胞因子对SD大鼠表皮干细胞分化为神经干细胞的影响

Effect of bFGF on the differentiation of epidermal stem cells to neural stem cells in SD rats

目的研究碱性成纤维细胞因子(bFGF)对大鼠表皮干细胞分化为神经细胞的影响。方法分离获得饲养了1～3天的新生SD大鼠表皮基底层组织,利用10 min快速贴壁法消化、分离获得表皮干细胞,于倒置显微镜下观察表皮干细胞的形态。培养表皮干细胞的培养基为K-SFM,然后按照不同密度表皮干细胞进行分组处理(0.1×10~7/mL,0.3×10~7/mL,0.5×10~7/mL,0.1×10~6/mL),每组加入20 ng/mL的bFGF,利用细胞免疫组织化学法检测细胞标志物Nestin和NSE的变化,以及观察细胞形态发生的变化。结果成功分离得到SD大鼠表皮干细胞;bFGF诱导后,0.3×10~7/mL组和0.1×10~7/mL组细胞第3天即可见细胞开始伸展生长,在约1周时细胞开始分化成神经细胞;且在细胞形态发生双极化改变的数目趋势上也具有一致性;Nestin和NSE检测均呈阳性表达。结论 bFGF有助于诱导表皮干细胞向神经细胞的分化,而且该诱导作用具有一定的细胞密度依赖性。

Objective To investigate the effect of basic fibroblast factor(bFGF)on the differentiation of epidermal stem cells into nerve cells in rats.Methods The epidermal basal layer tissue of newborn SD rats(1-3 day)were isolated and obtained.Epidermal stem cells were digested and isolatedby rapid attachment to a substrate,and the morphology of epidermal stem cells were observed under an inverted microscope.Epidermal stem cells were cultured with K-SFM culture medium.The epidermal stem cells were grouped according to the density as 0.1×10^7/mL,0.3×10^7/mL,0.5×10^7/mL,0.1×10^6/mL,and each group was added bFGF(20 ng/mL).The changes of cell markers Nestin and NSE were detected by immunohistochemistry,and the changes of cell morphology were observed.Results The epidermal stem cells of SD rat were isolated successfully.After bFGF induction,the 0.3×10^7/mL and 0.1×10^7/mL groups began to stretch growth in the third days,and the cells began to differentiate into nerve cells in about a week.The trend of the number of double polarization changes in the cell morphology was also consistent Nestin and NSE was positively expressed in those cells.Conclusion bFGF was helpful to induce the differentiation of epidermal stem cells into nerve cells,and the induction effect was dependent on the cell density.