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目的利用免疫磁珠分离成人骨髓神经生长因子受体(nervegrowthfactorreceptor,NGFR)阳性细胞,获得同质性骨髓基质干细胞(bonemarrowstromalcells,BMSCs)。方法采用Percoll密度梯度离心法分离成人骨髓中单个核细胞(mononuclearcells,MNCs)。对MNCs进行常规贴壁培养或应用磁分离技术分离NGFR+细胞。分别检测NGFR+细胞和常规贴壁培养所获BMSCs体外扩增和集落形成能力,分析其细胞表型和细胞周期,并进行成骨、成脂肪诱导。结果免疫磁珠分离获得NGFR+细胞的纯度为(90.4±4.7)%,NGFR+细胞较贴壁培养获得BMSCs具备更强增殖能力和成骨及成脂肪分化潜能。结论利用免疫磁珠分离骨髓NGFR+细胞可以获得同质性原始BMSCs。
OBJECTIVE: To isolate bone marrow stromal cells (BMSCs) from bone marrow cells of adult bone marrow using immunomagnetic beads (NGFR) -positive cells. Methods Mononuclear cells (MNCs) from human bone marrow were isolated by Percoll density gradient centrifugation. Conventional adherent culture of MNCs or isolation of NGFR + cells using magnetic separation techniques. The proliferation and colony formation ability of BMSCs from NGFR + cells and conventional adherent culture were detected respectively. The phenotype and cell cycle of BMSCs were detected, and osteogenic and adipogenic induction were performed. Results The purity of NGFR + cells isolated by immunomagnetic beads was (90.4 ± 4.7)%. NGFR + cells had stronger proliferative ability and osteogenic and adipogenic potential than BMSCs. Conclusion Isolation of bone marrow NGFR + cells by immunomagnetic beads can obtain homogeneous primary BMSCs.