bFGF对大鼠骨髓间充质干细胞向睾丸间质细胞分化的作用研究

发布时间：2018-04-12 12:42

本文选题：骨髓间充质干细胞 + 睾丸间质细胞　；参考：《山西医科大学》2015年硕士论文

【摘要】：目的:通过研究b FGF在骨髓间充质干细胞(BMSCs)定向分化为睾丸间质细胞(Leydigs)过程中的作用,探讨骨髓间充质干细胞(BMSCs)定向分化为睾丸间质细胞(Leydigs)的适宜方法。方法:通过全骨髓贴壁法获得大鼠骨髓间充质干细胞,将经纯化鉴定的BMSCs第3代细胞接种于六孔板,随机分为普通培养基对照组(A)、HCG+PDGF诱导组(B)、HCG+PDGF+5.0ng/ml b FGF诱导组(C)、HCG+PDGF+10.0ng/ml b FGF诱导组(D)、HCG+PDGF+20.0ng/ml b FGF诱导组(E),每组设3个复孔进行诱导,分别于第7天、14天、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)免疫荧光表达的阳性细胞数及其比例进行检测。结果:1)B、C、D、E组细胞诱导后,光学显微镜下细胞体积变大,细胞间互相连接呈长梭形或不规则形,贴壁生长,细胞核大,较圆,符合Leydig细胞的特点。2)随着诱导时间的增加、随着b FGF浓度的增加,B、C、D、E组睾酮水平逐渐升高,免疫荧光检测3β-HSD阳性表达细胞逐渐增多。其中:C、D、E组与B组,D、E组与C组间结果差异有统计学意义(P0.05);D、E组之间结果差异无统计学意义(P0.05)。诱导过程A组细胞培养液中未检测到睾酮,免疫荧光检测未见3β-HSD阳性细胞。结论:1)b FGF在大鼠BMSC细胞向Leydig细胞体外诱导分化过程中有明显促进作用。2)b FGF的浓度和诱导时间与其在大鼠BMSC细胞向Leydig细胞体外分化过程中的作用明显相关。
[Abstract]:Objective: to investigate the role of b FGF in the differentiation of bone marrow mesenchymal stem cells (BMSCs) into Leydigs (Leydigs), and to explore the appropriate method for the differentiation of BMSCs into Leydigs.Methods: rat bone marrow mesenchymal stem cells were obtained by whole bone marrow adherent method. The third generation of purified BMSCs cells were inoculated into the six-well plate.The control group was randomly divided into two groups: control group (control group), HCG PDGF PDGF 5.0ng/ml b FGF induction group, FGF induction group, HCG PDGF 10.0ng/ml b FGF induction group, HCG PDGF 20.0ng/ml b FGF induction group, each group was induced by 3 multiple holes, and morphological observation was made on the 7th day, 14 days and 21 days, respectively.The levels of testosterone and the number and proportion of 3 尾 -hydroxysteroid dehydrogenase 3 尾 -HSD-positive cells were detected.Results after induction, the volume of cells increased under optical microscope, the intercellular connections were fusiform or irregular, the cells adhered to the wall, and the nucleus was large and round, which accorded with the characteristics of Leydig cells. 2) with the increase of induction time.With the increase of the concentration of b FGF, the testosterone level and the expression of 3 尾 -HSD positive cells increased gradually.There was no significant difference in the results between group E and group B between group E and group C. There was no significant difference in results between group E and group C (P 0.05), and there was no significant difference between group D (P 0.05) and group C (P 0.05).No testosterone was detected in the culture medium of group A cells, but no 3 尾 -HSD positive cells were detected by immunofluorescence.Conclusion the concentration and induction time of FGF in the process of inducing the differentiation of rat BMSC cells into Leydig cells are significantly related to the differentiation of rat BMSC cells into Leydig cells in vitro.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R588

【参考文献】