碱性成纤维细胞生长因子对大鼠胰腺干细胞增殖的影响

Effect of base fibroblast growth factor on proliferation of rat pancreatic stem cells

目的探讨碱性成纤维细胞生长因子(bFGF)对大鼠胰腺干细胞增殖的影响。方法大鼠胰腺干细胞经体外纯化、扩增,观察其生长特性,免疫组化法检测CK-19和PDX-1的表达。研究不同浓度的bFGF对大鼠胰腺干细胞生长曲线的影响。结果原代培养的细胞经差速贴壁后,得到纯化的胰腺干细胞。免疫组化显示大鼠胰腺干细胞呈CK-19阳性和PDX-1阴性。对照组(无bFGF)、bFGF浓度分别为5,10,20 ng/ml各组的细胞数分别是3.29×105/ml,3.47×105/ml,3.83×105/ml和3.83×105/ml。加bFGF 3组的细胞数与对照组比较均有显著性差异(均P<0.01),10,20 ng/ml bFGF组细胞数与5 ng/ml bFGF组细胞数比较差异有显著性(P<0.01);10 ng/ml和20 ng/ml bFGF组差异无显著性(P>0.05)。结论5,10,20 ng/ml浓度的bFGF可明显促进SD大鼠胰腺干细胞增殖。

Objective To investigate the effect of basic fibroblast growth factor （bFGF） on proliferation of pancreatic stem cell （PSC） of SD rat. Methods PSCs were eultured in vitro after being purified. CK- 19 and Pdx- 1 expression were detected by inmaunohistochemistry to identify the proliferative characteristics of PSC. Different concentrations of bFGF were used to study the effect on grow curve of PSCs. Results The purified primary- cultured PSCs were obtained after repeated adherence. Immunohistochemistry results showed positive expression of CK - 19, and negative expression of Pdx - 1 in PSCs. The PSC number of control group,being cultured with no bFGF and of the experimental groups, being cultured with 5 ng/ml, 10 ng/ml and 20 ng/ml bFGF was 3.29×10^5/ml,3.47×10^5/ml,3.83×10^5/ml and 3.83 ×10^5/ml respectively. The PSC numbers of group cultured with 10 ng/ml and 20 ng/ml bFGF were significantly higher than that of 5 ng/ml bFGF group （ P 〈 0.01）. The cell numbers of all three bFGF groups were significantly higher than that of control group （no bFGF） （ P 〈 0.01 ）.Conclusion All three different bFGF concentrations （5 ng/ml, 10 ng/ml and 20 ng/ml） can significantly stimulate the proliferation of PSC.