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目的观察人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUC-MSCs)在体外向脂肪细胞诱导分化过程中细胞形态及表面抗原表达量变化。方法从脐带中分离培养间充质干细胞,至第3代,流式细胞仪检查其细胞表面抗原的表达量,加入成脂肪诱导剂体外诱导分化。诱导7 d,14 d,21 d,分别采用流式细胞仪分析细胞表面抗原的表达情况,结合油红O染色,观察细胞诱导分化情况。结果细胞培养至第3代,细胞表面表达CD90、CD44、CD29、CD71、CD73、CD105及CD13,不表达CD34、CD14、CD45、CD133、CD80、CD83、CD86、HLA-DR。成脂诱导7 d,细胞表面CD71表达量明显下降(P<0.05),但其他一些重要的标志物仍处于高表达水平;诱导14 d细胞表面抗原的表达量下降更为明显,如CD44为65.21%,CD71为42.05%(P<0.01),油红O染色有明显脂滴形成;诱导21 d细胞表面抗原的表达量均明显降低(P<0.01),油红O染色胞浆中充满红色油滴。结论 HUC-MSCs成脂肪诱导剂作用下,可诱导成脂肪细胞,且细胞表面抗原的表达量会在诱导过程中发生相应的改变。
Objective To observe the changes of cell morphology and surface antigen expression of human umbilical cord mesenchymal stem cells (HUC-MSCs) induced in vitro during adipocyte differentiation. METHODS: Mesenchymal stem cells (MSCs) were isolated and cultured from umbilical cord. The expression of cell surface antigens was detected by flow cytometry (FCM). The cells were induced to differentiate by adding adipogenic induction agent. Induction of 7 d, 14 d, 21 d, respectively, by flow cytometry analysis of cell surface antigen expression, combined with oil red O staining, observation of cell differentiation. Results The cells were passaged to the 3rd passage. CD90, CD44, CD29, CD71, CD73, CD105 and CD13 were expressed on the surface of the cells without expressing CD34, CD14, CD45, CD133, CD80, CD83, CD86 and HLA-DR. After 7 days of adipogenic induction, the expression of CD71 on the cell surface was significantly decreased (P <0.05), but some other important markers were still at high expression levels. The decreased expression of cell surface antigens on the 14th day was more obvious, for example, CD44 was 65.21 %, CD71 was 42.05% (P <0.01), and lipid droplets were formed by oil red O staining. The expression of cell surface antigens was significantly decreased on the 21st day after induction (P <0.01). The oil red O staining cytoplasm was filled with red oil drop. Conclusion HUC-MSCs can be induced into adipocytes under the action of fat-inducing agents, and the expression of cell surface antigens will be changed during the induction process.