胎儿肌肉来源细胞的分离及其成肌分化研究

发布时间：2018-04-27 03:14

本文选题：胎儿肌肉来源细胞 + Duchenne型肌营养不良　；参考：《中国现代医学杂志》2015年14期

【摘要】：目的观察胎儿肌肉来源细胞(f MDC)体外成肌分化能力以及在mdx鼠体内再生抗肌营养不良蛋白。方法使用含10%新生牛血清的DMEM/F12培养液,采用预贴壁方法从胎儿肌肉组织中分离f MDC。采用免疫染色鉴定肌肉特异标志的表达。肌内注射f MDC到6.5 Gy照射3 d后的mdx小鼠股直肌,1个月后采用免疫荧光染色法和RT-PCR方法检测人抗肌营养不良蛋白和基因的表达。结果采用预贴壁方法可从胎儿肌肉组织中分离到贴壁生长、梭型的f MDC。可在f MDC的一些长纤维细胞包浆中检测到MHC的表达,在一些细胞胞核中检测到成肌素的表达。可在注射f MDC的mdx小鼠肌肉中检测到人抗肌营养不良蛋白和基因的表达。结论 f MDC中存在MHC和成肌素的阳性表达,注射f MDC到照射过的mdx小鼠体内可以再生抗肌营养不良蛋白表达。
[Abstract]:Objective to observe the ability of myogenic differentiation of fetal muscle-derived cells (FMC) in vitro and the regeneration of anti-muscular dystrophy protein (AMP) in mdx mice. Methods DMEM/F12 medium containing 10% newborn bovine serum was used to isolate fMDC from fetal muscle tissue by pre-adherent method. The expression of muscle specific markers was identified by immunostaining. After intramuscular injection of f MDC to 6.5 Gy for 3 days, the expression of human anti-muscular dystrophy protein and gene was detected by immunofluorescence staining and RT-PCR method. Results the adherent and fusiform f MDCs could be isolated from fetal muscle tissue by pre adherent method. The expression of MHC was detected in the long fibroblast envelope of f MDC, and the expression of sarcoblast was detected in the nucleus of some cells. Human anti-muscular dystrophy protein and gene expression were detected in mdx mice injected with f MDC. Conclusion there is positive expression of MHC and creatinin in f MDC, and the expression of anti muscular dystrophy protein can be regenerated by injecting f MDC into irradiated mdx mice.
【作者单位】：成都军区昆明总医院云南省干细胞工程实验室;昆明医科大学昆明总医院临床学院;
【基金】：云南省科技创新人才计划(No:2011HB050) 云南省科技创新平台建设计划(No:2013DA004)
【分类号】：R714.5

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