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目的:对高原红细胞增多症(HAPC)患者骨髓间充质干细胞(MSCs)进行分离培养,为进一步探讨其生物学特性奠定基础。方法:采用密度梯度离心和贴壁筛选法对10例HAPC和10例正常人骨髓MSCs进行分离培养,倒置显微镜下观察MSCs形态,流式细胞仪行细胞表面抗原检测,检测MSCs生长情况,判断其增殖能力。结果:骨髓密度梯度离心法分离骨髓单个核细胞(BMMNC)后培养,骨髓MSC均分离培养成功,贴壁细胞呈梭形;两组骨髓MSCs均表达CD29、CD105和CD13,不表达CD45、CD4、CD8、CD14、CD3、CD34和HLA-DR(P>0.05)。HAPC骨髓MSCs增殖能力高于正常骨髓(P<0.01)。结论:HAPC患者骨髓MSCs体外可有效分离培养,所培养的细胞成分单一,可供进一步实验应用。
OBJECTIVE: To isolate and culture bone marrow mesenchymal stem cells (MSCs) from patients with high altitude polycythemia (HAPC), and to lay the foundation for further exploration of their biological characteristics. Methods: MSCs were isolated from 10 HAPC and 10 normal human bone marrow using density gradient centrifugation and adherence screening. Morphological changes of MSCs were observed under inverted microscope. Cell surface antigens were detected by flow cytometry. Proliferative ability. Results: Bone marrow mononuclear cells (BMMNCs) were isolated by bone marrow density gradient centrifugation. The bone marrow MSCs were isolated and cultured successfully, and the adherent cells were fusiform. MSCs of both groups expressed CD29, CD105 and CD13, but did not express CD45, CD4, CD8, CD14, CD3, CD34 and HLA-DR (P> 0.05). The proliferation of HAPC bone marrow MSCs was higher than that of normal bone marrow (P <0.01). Conclusion: The bone marrow MSCs of HAPC patients can be effectively isolated and cultured in vitro, and the cultured cells have a single component for further experimental application.