本文選題：骨髓間充質(zhì)干細胞 + 睪丸間質(zhì)細胞　； 參考：《山西醫(yī)科大學》2015年碩士論文

【摘要】：目的:通過研究b FGF在骨髓間充質(zhì)干細胞(BMSCs)定向分化為睪丸間質(zhì)細胞(Leydigs)過程中的作用,探討骨髓間充質(zhì)干細胞(BMSCs)定向分化為睪丸間質(zhì)細胞(Leydigs)的適宜方法。方法:通過全骨髓貼壁法獲得大鼠骨髓間充質(zhì)干細胞,將經(jīng)純化鑒定的BMSCs第3代細胞接種于六孔板,隨機分為普通培養(yǎng)基對照組(A)、HCG+PDGF誘導組(B)、HCG+PDGF+5.0ng/ml b FGF誘導組(C)、HCG+PDGF+10.0ng/ml b FGF誘導組(D)、HCG+PDGF+20.0ng/ml b FGF誘導組(E),每組設3個復孔進行誘導,分別于第7天、14天、21天進行形態(tài)學觀察、培養(yǎng)液睪酮水平及3β-羥化類固醇脫氫酶(3β-HSD)免疫熒光表達的陽性細胞數(shù)及其比例進行檢測。結(jié)果:1)B、C、D、E組細胞誘導后,光學顯微鏡下細胞體積變大,細胞間互相連接呈長梭形或不規(guī)則形,貼壁生長,細胞核大,較圓,符合Leydig細胞的特點。2)隨著誘導時間的增加、隨著b FGF濃度的增加,B、C、D、E組睪酮水平逐漸升高,免疫熒光檢測3β-HSD陽性表達細胞逐漸增多。其中:C、D、E組與B組,D、E組與C組間結(jié)果差異有統(tǒng)計學意義(P0.05);D、E組之間結(jié)果差異無統(tǒng)計學意義(P0.05)。誘導過程A組細胞培養(yǎng)液中未檢測到睪酮,免疫熒光檢測未見3β-HSD陽性細胞。結(jié)論:1)b FGF在大鼠BMSC細胞向Leydig細胞體外誘導分化過程中有明顯促進作用。2)b FGF的濃度和誘導時間與其在大鼠BMSC細胞向Leydig細胞體外分化過程中的作用明顯相關(guān)。
[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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R588

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