大鼠应激耐受多系分化细胞的分离、扩增改良技术

Isolation and amplified culture of multilineage differentiating stress enduring cells

目的探讨体外从大鼠骨髓间充质干细胞(BMSCs)中分离应激耐受多系分化细胞(Muse cells)及其扩增培养的技术方法。方法分离、扩增、鉴定BMSCs,应用长时间应激方法酶消化分离Muse细胞,悬浮扩增培养4d后Western blot鉴定特异性抗原。结果第3代生长良好的BMSCs中表达CD29、CD90而不表达CD45的占细胞总数的97.89%。长时间酶消化结合悬浮培养后,以酶消化8h组细胞表达CD105显著高于其余各组(P<0.05),符合Muse细胞特征。结论 Muse细胞可由BMSCs经消化分离扩增获得,可望为其进一步研究应用奠定基础。

Objective To investigate the isolation and amplified culture method of the multilineage differentiating stress enduring cells （Muse cells）. Methods Rat bone marrow mesenchymal stem cells （BMSCs） were isolated, amplified and identified. Muse cells were isolated by long-term trypsin incubation from cultured BMSCs; then they were suspension cultured for 4 days and identified by western blot. Results 97.89 % of the third-passage cells were positive for CD29, CD90 and negative for CD45 on flow cytometry. After long-term trypsin incubation and suspension culture, western blot detected the specific marker of Muse cells. The result indicated that the CD105 expression of cells after 8-hour trypsin incubation and suspension culture was significantly higher than that in the other groups （P〈0.05）. Conclusion Muse cells can be isolated and amplified from BMSCs.