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目的观察不同浓度碱性成纤维细胞生长因子(bFGF)对体外培养人羊膜间充质干细胞(HAMSCs)增殖的影响,以探索体外培养HAMSCs的最适条件。方法分离、培养HAMSCs,选取第3代细胞进行鉴定及检测。实验组将bFGF分为0.1、0.5、1、5、10、20、40、80ng/ml共8个浓度组;阴性对照组只加HAMSCs,不加bFGF;空白对照组只加培养基。各组孵育48h后加入Am-Blue试剂,继续孵育,待培养基由靛青蓝色开始变成粉红色时取出用酶标仪测量各组的吸光度(OD),重复3次实验。结果 0.5、1、5、10、20、40、80ng/ml浓度组与对照组相比,差异均有统计学意义(P<0.05);10ng/ml组与0.5、1、5ng/ml组相比,差异均有统计学意义(P<0.05);10ng/ml组与40、80ng/ml组相比,差异无统计学意义(P=0.067),表明bFGF促增殖作用最强的最小浓度为10ng/ml。结论 bFGF能够促进HAMSCs增殖,且促增殖作用最强的最小浓度为10ng/ml。
Objective To observe the effects of different concentrations of basic fibroblast growth factor (bFGF) on the proliferation of cultured human amniotic mesenchymal stem cells (HAMSCs) in vitro to explore the optimal conditions for culturing HAMSCs in vitro. Methods HAMSCs were isolated and cultured, and the third generation of cells were selected for identification and detection. The experimental group bFGF divided into 0.1,0.5,1,5,10,20,40,80 ng / ml a total of eight concentration groups; negative control group only HAMSCs, without bFGF; blank control group only with medium. After 48 hours of incubation, Am-Blue reagent was added and the incubation was continued. When the medium began to turn pink from indigo blue, the absorbance (OD) of each group was measured by microplate reader and the experiment was repeated 3 times. Results Compared with the control group, the differences were statistically significant (P <0.05), while those of 0.5ng / ml, 0.5ng, 10ng, 40ng / (P <0.05). There was no significant difference between 10ng / ml group and 40,80ng / ml group (P = 0.067), which indicated that the strongest promoting effect of bFGF was 10ng / ml. Conclusions bFGF can promote the proliferation of HAMSCs, and the strongest promoting effect of proliferation is 10ng / ml.