胎儿肌肉来源细胞的分离及其成肌分化研究  

Isolation and myogenic differentiation of fetal muscle-derived cells

作　　者：庞荣清[1] 李自安[1] 王强[1] 杨建勇[1] 朱慧[2] 何洁[1] 朱向情[1] 

机构地区：[1]成都军区昆明总医院云南省干细胞工程实验室,云南昆明650032 [2]昆明医科大学昆明总医院临床学院,云南昆明650031

出　　处：《中国现代医学杂志》2015年第14期21-24,共4页China Journal of Modern Medicine

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)

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。[Objective] To investigate in vitro myogenic differentiation capacity of fetal muscle-derived cells （fMDC） and regeneration of dystrophin in mdx mice. [Methods ] FMDC were isolated from fetal muscle tissue by using preplate method in DMEM/F12 medium containing 10% newborn calf serum. Muscle-specific markers were identified by immunostaining in fMDC. One month later, human dystrophin protein expression was detected by immunofluorescence staining while dystrophin mRNA expression was detected by RT-PCR in the rectus femoris of mdx mice that received intramuscular injection of fMDC 3 days after irradiation at 6.5 Gy. [Results] FMDC with adherent growth and spindle shape were isolated from fetal muscle by preplate method. Myosin heavy chain （MHC） expression in cytoplasm was found in some long fiber of fMDC while myogenin expression was detected in the nuclei of some fMDC. Human dystrophin protein and gene expres- sions were detected in the muscle of irradiated mdx mice injected with fMDC. [ Conclusion] Positive expres- sions of MHC and myogenin are present in fMDC, and regeneration of dystrophin could be detected in the muscle of irradiated mdx mice with fMDC injection.

关 键 词：胎儿肌肉来源细胞 DUCHENNE型肌营养不良 动物模型 抗肌营养不良蛋白 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]