摘要
目的:探讨脂肪组织源性间充质干细胞体外分离培养的方法并将其向心肌细胞方向诱导分化。方法:无菌条件下获取2月龄SD大鼠腹部肠系膜脂肪组织,0.08%胰酶消化分离出单个核细胞,接种7～10d后形成单层贴壁长梭形细胞。0.25%胰酶消化传代,第3代细胞用于鉴定细胞表面分子及诱导分化。浓度为10μmol/L5-氮胞苷与分离细胞共同培养24h,3周后应用抗desmin及抗α-actnin抗体对诱导细胞进行免疫化学鉴定。结果:所分离细胞约90%表达间充质干细胞表面抗原CD13、CD44。5-氮胞苷诱导后3周,所培养的细胞形成desmin及α-actnin染色阳性的心肌细胞团。结论:体外通过消化法可从脂肪组织中分离出大量间充质干细胞,这些细胞经5-氮胞苷诱导后分化为心肌细胞。
Objective： To establish a method for isolating and culturing the mesenchymal stem cells derived from adipose tissue in vitro and inducing them into cardiomyocytes in rats. Methods： The adipose tissue was obtained from the mesentery of the two-month SD rats. 0.08% trypsin was used to digest the adipose tissue and obtain the mesenchymal stem cells. The shuttle-like monolayer cells were obtained 7～10 days after seeded. The third passage cells were used to induce into cardiomyocytes and detect the surface molecule. The cells were cultured with 10μmol/L 5-AZA for 24 hours. The anti-desmin and anti-α-actnin monoclonal antibodies were used to detect the induced cells after 3 weeks by immunocytochemistry method. Results： 90% cells were CD13, CD44 positive mesenchymal stem cells. After induced with 5-AZA for 3 weeks, the cells could differentiate into desmin and α-actnin positive cardiomyocytes group. Conclusion： Plenty of mesenchymal stem cells can be obtained in vitro using digestion method, and these cells can differentiate into cardiomyocytes induced by 5-AZA.
出处
《山东大学学报（医学版）》
CAS
北大核心
2005年第7期578-581,共4页
Journal of Shandong University:Health Sciences
