CAJ | 학술논문

目的探讨5-氮杂脱氧胞苷(5-AzaC)体外诱导人羊水来源间充质干细胞向成肌细胞样细胞分化的可行性.方法 B超引导下穿刺抽得孕中期羊水,体外分离、培养得到羊水来源间充质干细胞,传代后计数细胞并绘制生长曲线.体外成骨、成脂诱导观察人羊水来源间充质干细胞多向分化能力.采用5-AzaC对羊水来源间充质干细胞成肌诱导2周,观察细胞形态学变化,RT-PCR、免疫荧光染色方法鉴定成肌细胞特异mRNA及蛋白表达.结果人羊水来源间充质干细胞体外培养后迅速进入对数生长期,培养7 d未达到平台期.茜素红和油红O染色证实人羊水来源间充质干细胞可诱导分化为成骨、成脂细胞样细胞.人羊水来源间充质干细胞经5-AzaC诱导2周逐渐变长梭形.而对照组细胞呈扁平多角形.免疫荧光染色及RT-PCR结果提示实验组细胞表达肌结蛋白(Desmin)、肌钙蛋白I(Tn I)、横纹肌辅肌动蛋白(α-Actinin)及mRNA;对照组呈阴性.结论人羊水来源间充质干细胞体外增殖能力强,能分化为成骨、成脂细胞样细胞.5-AzaC能在体外诱导人羊水来源间充质干细胞向成肌细胞样细胞分化.
목적 탐토5-담잡탈양포감(5-AzaC)체외유도인양수래원간충질간세포향성기세포양세포분화적가행성.방법 B초인도하천자추득잉중기양수,체외분리、배양득도양수래원간충질간세포,전대후계수세포병회제생장곡선.체외성골、성지유도관찰인양수래원간충질간세포다향분화능력.채용5-AzaC대양수래원간충질간세포성기유도2주,관찰세포형태학변화,RT-PCR、면역형광염색방법감정성기세포특이mRNA급단백표체.결과 인양수래원간충질간세포체외배양후신속진입대수생장기,배양7 d미체도평태기.천소홍화유홍O염색증실인양수래원간충질간세포가유도분화위성골、성지세포양세포.인양수래원간충질간세포경5-AzaC유도2주축점변장사형.이대조조세포정편평다각형.면역형광염색급RT-PCR결과제시실험조세포표체기결단백(Desmin)、기개단백I(Tn I)、횡문기보기동단백(α-Actinin)급mRNA;대조조정음성.결론 인양수래원간충질간세포체외증식능력강,능분화위성골、성지세포양세포.5-AzaC능재체외유도인양수래원간충질간세포향성기세포양세포분화.
Objective To investigate the efficacy of 5-AzaC to induce the myogenic differentiation of human amniotic fluid-derived mesenchymal stem cells (HAFDMSCs) in vitro. Methods HAFDMSCs were isolated from second trimester amniotic fluid and cultured in vitro. Osteogenic and adipogenic differentiation was carried out to study multipotent potential of HAFDMSCs. HAFDMSCs were cultured in myogenic differentiation medium temporarily supplemented by 10 ummol/L 5-AzaC for 24hours or complete medium as experimental and control groups, respectively. The differentiated cells were identified morphologically and by the expressions of myocyte specific proteins after induction for 2 weeks. Results HAFDMSCs grew well in vitro and didn't reach the plateau phase even after being cultured for 7 days. Adipogenic and osteogenic differentiation potential of HAFDMSCs was confirmed by Alizarin red and oil red O staining. After 5-AzaC induction, HAFDMSCs appeared elongated shape.Immunofluorescence staining and RT-PCR confirmed the expression of myocyte specific phenotypes like Desmin,Tn I and α-Actinin. Conclusions 5-AzaC can induce the HAFDMSCs to differentiate into myoblast-like cells in vitro.