摘要
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 explore the role of the basic fibroblast growth factor（b FGF） in the directional differentiation of bone marrow mesenchymal stem cells（ BMSCs） into Leydig cells. Methods： After purification and identification,we inoculated the thirdgeneration BMSCs of SD rats onto a six-orifice board and then randomly divided them into groups A（ normal saline control）,B（ human chorionic gonadotropin [h CG] ＋ platelet-derived growth factor [PDGF] induction）,C（ h CG ＋ PDGF ＋ 5. 0 ng / ml b FGF induction）,D（ h CG ＋ PDGF ＋ 10. 0 ng / ml b FGF induction）,and E（ h CG ＋ PDGF ＋ 20. 0 ng / ml b FGF induction）. On the 7th,14 th and 21 st day of induction,we observed the morphological changes of the cells and measured the level of testosterone（ T） and expression of 3 beta hydroxy steroid dehydrogenase（3β-HSD） in the supernatant by immunofluorescence staining. Results： After induction,the BMSCs of groups B,C,D,and E exhibited microscopic features of enlarged size,inter-connection,long-shuttle or irregular shape,adherent growth,and large round nuclei,all characteristic of Leydig cells. With the prolonging of time and enhanced concentration of b FGF,gradual increases were observed in the T level and the count of 3β-HSD-positive BMSCs in the four induction groups,with statistically significant differences between group B and groups C,D,and E（ P 〈 0. 05）,as well as between group C and groups D and E（ P 〈 0. 05）,but not between D and E（ P 〉 0. 05）. Conclusion： The b FGF has an obvious promoting effect in the in vitro induced differentiation of rat BMSCs into Leydig cells.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2015年第6期494-499,共6页
National Journal of Andrology
基金
山西省高校"131"领军人才工程项目~~
