骨髓间充质细胞与多系分化持续应激细胞的蛋白 质谱分析比较

目的：比较骨髓间充质细胞（hBMSCs）和多系分化持续应激细胞（Muse细胞）蛋白表达的差异。方法：通过密度梯度离心和差速贴壁法从健康人骨髓中分离得到hBMSCs，流式细胞仪检测其中SSEA-3/CD105双阳性细胞；利用长时间胰酶孵育法从hBMSCs中分离得到Muse细胞，利用qPCR技术在mRNA水平检测多能干细胞标记物的表达情况；提取hBMSCs和Muse细胞蛋白，应用iTRAQ标记结合online 2D LC/MS/MS蛋白组学技术定量评估蛋白质表达谱。结果：hBMSCs中SSEA-3和CD105双阳性细胞占0.51%；长时间胰酶孵育法可以成功分离出Muse细胞，悬浮可呈球状生长，且SSEA-3、Sox-2、Oct-4表达均高于hBMSCs；蛋白质谱共检测4 377个蛋白，相对于hBMSCs，Muse细胞有27个上调蛋白和68个下调蛋白。结论：hBMSCs和Muse细胞蛋白表达基本相同，仅有95种蛋白存在差异。

Comparison of bone marrow mesenchymal cells and multilineage differentiating stress enduring cells by protein spectrum analysis

Objective: To compare the differences in protein expression between hBMSCs and Muse cells. Methods:hBMSCs were isolated from human bone marrow by density gradient centrifugation and differential adherence method, and detected by flow cytometry. Muse cells were isolated from hBMSCs by long-term trypsin incubation to detect pluripotent stem cell characteristics. The hBMSCs and Muse cell proteins were extracted and analyzed. Results: SSEA-3 and CD105 double positive cells accounted for 0.51% in hBMSCs. Muse cells could be successfully isolated by long-term trypsin incubation. The suspension could grow in a spherical shape, and the expression of SSEA-3, Sox-2 and Oct-4 was high. A total of 4377 proteins were detected in the protein profiles of hBMSCs, with 27 up-regulated proteins and 68 down-regulated proteins relative to hBMSCs by 95% confidence interval Muse. Conclusion: The expression of hBMSCs and Muse cell proteins is basically the same, and only 95 proteins are different.