摘要
目的从基因角度求证脐带间充质干细胞(MSCs)神经巢蛋白nestin mRNA表达。方法取新鲜脐带组织行MSCs分离、培养及鉴定。采用胰酶消化法计数MSCs,流式细胞仪检测细胞活性,提取原代、一代、二代细胞的mRNA,采用实时荧光定量PCR法检测nestin mRNA表达。结果脐带中分离出的细胞贴壁生长状况较好,其中大部分细胞呈梭形生长,传代后细胞与原代细胞生长形态相似,生长加快;个别细胞呈神经样生长。MSCs呈nestin mRNA阳性表达,其表达量在传代后有明显减少;与nestin蛋白在神经前体细胞向神经细胞分化时的变化一致。结论脐带MSCs存在nestin mRNA表达;传代后nestin mRNA表达量虽减小,但仍可作为备选种子细胞进行移植。
Objective To observe the expression of nestin nestin mRNA in umbilical cord mesenchymal stem cells （MSCs） from the perspective of gene .Methods The fresh umbilical cord tissues underwent MSCc separation and then were cultured and purified .Cells from the umbilical cord were counted by enzymatic digestion , and the activity was detec-ted by flow cytometry .The primary, first-generation and second-generation cellular mRNA were extracted to detect the ex-pression of nestin mRNA by real-time PCR.Results The cells isolated from umbilical cord grew in good condition , in which a large part of the cells grew in fusiform , cells after passage had the similar morphology with primary cells and grew faster.There were individual cells which were inneural-like growth.Nestin mRNA expression was found in umbilical cord MSCs, and the expression level after the passage was significantly decreased , meanwhile, the nestin protein changes were consistent with that of differentiation from the neural precursor cells into neural cells .Conclusion The nestin mRNA ex-pression was found in the MSCs tissue genes .Even the expression level of nestin mRNA decreases after passage , it still can be used as an alternative seed cells for transplantation .
出处
《山东医药》
CAS
2014年第10期4-6,共3页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81360189)
关键词
间充质干细胞
脐带
神经巢蛋白
实时荧光定量聚合酶链反应
mesenchymal stem cells,umbilical cord
nestin
real-time fluorescence quantitative polymerase chain reaction
