脂肪间充质干细胞向睾丸间质细胞的诱导分化

Induced differentiation of adipose-derived mesenchymal stem cells into Leydig cells

背景:近年来干细胞的发展,为治疗雄激素缺乏疾病提供了新的思路。目的:探讨脂肪间充质干细胞是否可分化为睾丸间质细胞。方法:取生长状态良好的第3代大鼠脂肪间充质干细胞,分2组培养:实验组加入0.1 mg/L人绒毛膜促性腺激素、10.0μg/L血小板源性生长因子及10.0μg/L碱性成纤维细胞生长因子,进行向睾丸间质细胞的诱导培养;对照组不加入上述细胞生长因子,常规培养。诱导培养1,7,14,21 d,进行相关指标检测。结果与结论:(1)免疫荧光染色:对照组无3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehydrogenase,3β-HSD)阳性细胞;实验组可见3β-HSD阳性细胞,并且随着诱导时间的延长,3β-HSD阳性细胞逐渐增多,荧光染色增强;(2)睾酮分泌水平:对照组无睾酮分泌;诱导培养7 d后,实验组检测到睾酮分泌,并且随着诱导时间的延长,睾酮分泌增加;(3)RT-PCR检测:对照组无黄体生成素受体、类固醇合成急性调节蛋白、3β-HSD阳性条带;实验组诱导培养7 d后,可见黄体生成素受体、类固醇合成急性调节蛋白、3β-HSD阳性条带,并且随着诱导培养时间的延长,阳性条带增强;(4)结果表明,脂肪间充质干细胞可分化为睾丸间质细胞。

BACKGROUND:The recent development of stem cells has provided new ideas for the treatment of androgen-deficient diseases.OBJECTIVE:To investigate whether adipose-derived mesenchymal stem cells can differentiate into Leydig cells METHODS:Passage 3 rat adipose-derived mesenchymal stem cells that grew well were taken and cultured in the medium with(experimental)or without(control)0.1 mg/L human chorionic gonadotropin,10.0μg/L platelet-derived growth factor and 10.0μg/L basic fibroblast growth factor.Indicator detection was done at 1,7,14,24 days of induced culture.RESULTS AND CONCLUSION:(1)Immunofluorescence staining results showed that there were no 3β-hydroxysteroid dehydrogenase(3β-HSD)positive cells in the control group,while the number of 3β-HSD positive cells was gradually increased in the experimental group with the induction time,which presented with fluorescence enhancement.(2)There was no secretion of testosterone in the control group,while in the experimental group,testosterone secretion was detected at 7 days of induced culture,and moreover,the testosterone level was increased with the induction time.(3)RT-PCR findings showed no luteinizing hormone receptor,steroidogenic acute regulatory protein,and 3β-HSD positive bands in the control group,while these positive bands appeared in the experimental group after 1 day of induction,and strengthened with the induction time.To conclude,adipose-derived mesenchymal stem cells can be induced to differentiate into Leydig cells.