【作者】 张志远；

【导师】 孙杰；

【作者基本信息】 上海交通大学 ， 儿科学， 2016， 硕士

【摘要】 目的:探讨人脐带间充质干细胞(Human Umbilical Cord Mesenchymal Stem Cells,HUMSCs)在大鼠体内分化为Leydig细胞并治疗雄性性腺功能低下症的可行性。方法:采用贴壁法分离培养HUMSCs。通过腹腔注射二甲磺酸乙烷(ethane dimethane sulfonate,EDS)的方式建立雄性大鼠性腺功能低下模型,采用荧光染料氯甲基-1,1-十八烷基-3,3,3’,3’-四甲基-吲哚-羧花青-高氯酸盐(Chlor Methylbenzamido-1,1-Dioctadecyl-3,3,3’,3’-tetramethylin-docarbocyamine,CM-Dil)标记HUMSCs后将HUMSCs移植入模型大鼠睾丸间质内。三周后通过免疫组化与流式细胞术检测大鼠睾丸内HUMSCs细胞色素p450家族成员11A1(Cytochrome P450,Family 11,Subfamily A,Polypeptide 1,CYP11A1)与3β羟基类固醇脱氢酶(3β-Hydroxy Steroid Dehydrogenase,3β-HSD)的表达情况,并通过流式细胞术检测细胞分化效率。比较实验组与对照组大鼠血清睾酮水平,观察移植HUMSCs对动物雄激素恢复的治疗作用。结果:细胞移植三周后,部分CM-Dil标记的HUMSCs表达了CYP11A1,通过消化睾丸后流式分选CM-Dil阳性细胞并对其进行培养,发现这些细胞仍然能够在体外增殖,且这些CM-Dil阳性细胞中大约有24%可表达3β-HSD。血清睾酮检测发现HUMSCs移植后三周,治疗组血睾酮显著高于对照组。结论:HUMSCs可在睾丸内向Leydig细胞分化,并可显著提高实验动物的血清睾酮水平。更多还原

【Abstract】 OBJECTIVE: To explore the feasibility of Human Umbilical Cord Mesenchymal Stem Cells(HUMSCs)differentiation to Leydig cells in vivo and investigate the treatment effect of HUMSCs on male hypogonadism.METHODS: Using tissue attachment to culture HUMSCs,then transplant Chlor Methylbenzamido-1,1-Dioctadecyl-3,3,3’,3’-tetramethylin-docarbocyamine,(CM-Dil)stained HUMSCs in ethane dimethane sulfonate(EDS)treated rats.Detect Cytochrome P450,Family 11,Subfamily A,Polypeptide 1(CYP11A1)and 3β-Hydroxy Steroid Dehydrogenase(3β-HSD)by immunohistochemical and flowcytometry 3 weeks after tansplantation.Test the blood testosterone level to observe the medical effect of HUMSCs on hypogonadsim.RESULTS: 3 weeks after tansplantation,some of the CM-Dil stained HUMSCs expressed CYP11A1,CM-Dil stained HUMSCs are still able to proliferate in vitro after flow cytometry sorting,24% of these cells were postive for 3β-HSD test.The blood testosterone level of Experiment group is significantly higher than control group.CONCLUSION: HUMSCs are able to differentiate to Leydig cells in vivo,and HUMSCs can help the recovery of rats’ blood testosterone.更多还原