摘要：目的 研究成人骨髓间充质干细胞(MSCs)在诱导条件下定向分化为成骨细胞过程中,细胞表面分子的表达及变化特征。方法 从人骨髓中分离有核细胞,在含有15％胎牛血清、维生素C和β-甘油磷酸钠的α-MEM培养基中培养。在培养早期加入10-8mol/L地塞米松 (实验组)或保持基础培养状态（对照组1）,同时部分细胞仅培养于α-MEM培养基中(对照组2)。于培养第7、12、17天分别收集实验组和各对照组细胞,用流式细胞方法观察细胞表面分子CD45、CD34、CD117、CD90和人白细胞抗原-DR（HLA-DR）的表达,并定量对比分析。结果 成人骨髓MSCs在体外诱导条件下分化为具有成骨细胞特征的成熟细胞。培养第7天实验组、对照组1和对照组2均少量表达CD45,第12、17天CD45表达转为阴性。各组各时间点CD34、CD117表达均为阴性。培养第12天,实验组和各对照组细胞CD90表达均较第7天升高,以对照组升高明显。实验组细胞HLA-DR表达随着成骨细胞分化成熟逐渐上升,而对照组细胞HLA-DR表达下降。结论 MSCs在诱导条件下可以向成骨细胞分化。细胞表面分子的表达在细胞分化过程中呈特征性改变,是成骨细胞分化早期的一个重要标志。
Abstract
ABSTRACT:Objective To explore the phenotypic changes of some cell surface antigens in the process that the bone marrow-derived mesenchymal stem cells (MSCs) differentiate into osteoblast lineage under certain conditions. Methods The mononuclear cells were isolated from human bone marrow and cultured in the medium in the presence (experimental group) or absence (control group 1) of dexamethasone, ascorbic acid and β-glycerophosphate, or in the α-MEM only (control group 2). The expressions of CD45, CD34, CD117, CD90,and HLA-DR were examined by flow cytometry on culture day 7, 12, and 17. Results Human bone marrow MSCs can differentiate into mature cells with the characteristics of osteoblasts in vitro. The expression of CD45 was low in the experimental group and in the control groups on culture day 7, and became negative from day 12 in all groups. The expressions of CD34 and CD117 were negative at all time points. The expression of CD90 increased on day 12 in all groups, and was more obvious in the control groups. The expression of HLA-DR was gradually elevated along with the differentiation and maturation of osteoblasts in the experimental group, but dropped in the control groups in the later time points. Conclusions MSCs can differentiate into mature osteoblasts after induction. The expression of cell surface antigens during differentiation has characteristic changes, which may be key markers in the early stage of osteoblasts differentiation.
关键词
骨髓间充质干细胞 /
成骨细胞 /
细胞表面分子
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Key words
bone marrow-derived mesencdymal stem cells /
osteoblast /
cell surface anticens
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参考文献
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