bfgf对大鼠骨髓间充质干细胞向睾丸间质细胞分化的作用分析word格式论文.docx

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 中 文 摘 要目的：通过研究 bFGF 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 bFGF 诱导组(C）、HCG+PDGF+10.0ng/ml bFGF 诱导组（D）、 HCG+PDGF+20.0ng/ml bFGF 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 bFGF 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：I1）bFGF 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）bFGF 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。关键词：骨髓间充质干细胞；睾丸间质细胞；碱性成纤维细胞生长因子；迟发型性 腺功能减退IIThe role of bFGF on the differentiation from rat bone marrow mesenchymal stem cells to the leydig cellsAbstractObjective:Through examining the roles of bFGF in the directional differentiation of bone marrow mesenchymal stem cells (BMSCs) into leydig cells (Leydigs), this paper explores appropriate approaches by which BMSCs could be directionally differentiated into Leydigs.Methods:The third-generation BMSCs of rats with purification appraisal were inoculated onto a six-orifice board, and then were randomly divided into control group (A), HCG + PDGF induction group (B), HCG + PDGF + 5.0 ng/ml bFGF induction group (C), HCG + PDGF + 10.0 ng/ml bFGF induction group (D) and HCG + PDGF + 20.0 ng/ml bFGF induction group (E). On the 7th, 14th and 21st day of the research, morphological observation, culture of testosterone level and immunofluorescence test for 3 beta hydroxy steroid dehydrogenase (3 beta HSD)were conducted respectively.Results:1) Cells of B, C, D and E group after induction indicated the features of microscopic were bigger size, being connected with each other in long shuttle or irregular shape, adherent growing next to walls, and having larger an