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目的 :研究碱性成纤维细胞生长因子(b FGF)在大鼠骨髓间充质干细胞(BMSCs)定向分化为睾丸Leydig细胞过程中的作用,探讨BMSCs定向分化为Leydig细胞的适宜方法。方法:将经纯化鉴定的SD大鼠BMSCs第3代细胞接种于6孔板,随机分为对照组(A)、人绒毛膜促性腺素(h CG)+血小板源性生长因子(PDGF)诱导组(B)、h CG+PDGF+5.0 ng/ml b FGF诱导组(C)、h CG+PDGF+10.0 ng/ml b FGF诱导组(D)、h CG+PDGF+20.0 ng/ml b FGF诱导组(E),分别于第7、14、21天进行形态学观察、培养液睾酮水平及3β-羟化类固醇脱氢酶(3β-HSD)免疫荧光检测。结果:B、C、D、E组细胞诱导后体积变大,互相连接呈长梭形或不规则形,贴壁生长,核大、较圆,符合Leydig细胞的特点。随着时间和b FGF浓度的增加,B、C、D、E组睾酮水平逐渐升高,3β-HSD阳性表达细胞逐渐增多,C、D、E组与B组,D、E组与C组间差异有统计学意义(P均<0.05);D、E组之间差异无统计学意义(P>0.05)。结论:b FGF在大鼠BMSCs向Leydig细胞体外诱导分化过程中有明显促进作用。
Objective: To investigate the role of basic fibroblast growth factor (b FGF) in the directional differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into testicular Leydig cells and to explore the suitable method for the directional differentiation of BMSCs into Leydig cells. Methods: The third generation of purified SD rat BMSCs were seeded on a 6-well plate and randomly divided into control group (A), human chorionic gonadotropin (h CG) and platelet-derived growth factor (PDGF) (C), h CG + PDGF + 10.0 ng / ml b FGF induction group (D), h CG + PDGF + 20.0 ng / ml b FGF (E) group. Morphological observation was performed on the 7th, 14th and 21st day respectively. The level of testosterone in the culture medium and the immunofluorescence of 3β-hydroxysteroid dehydrogenase (3β-HSD) were detected. Results: After induction, the volume of cells in groups B, C, D and E became larger and became fusiform or irregular in shape, adherently growing, bigger in nucleus and larger in circle, which accorded with the characteristics of Leydig cells. The testosterone levels in groups B, C, D and E gradually increased with the increase of time and b FGF concentration, and the number of 3β-HSD positive cells gradually increased. The number of cells in groups C, D, E and B, D, E and C There was significant difference between the two groups (P <0.05). There was no significant difference between D and E groups (P> 0.05). Conclusion: b FGF promotes the differentiation of BMSCs into Leydig cells in vitro.