血管内皮生长因子C诱导骨髓间充质干细胞分化为淋巴管内皮细胞的研究

Vascular endothelial growth factor C induces differentiation of bone marrow mesenchymal stem cells into lymphatic endothelial cells

目的研究骨髓间充质干细胞向淋巴管内皮细胞分化的潜能及条件,为淋巴管再生和重建提供理想的细胞来源。方法分离SD大鼠骨髓间充质干细胞,流式细胞仪检测骨髓间充质干细胞表面抗原CD31、CD14、CD29和CD90。将纯化的骨髓间充质干细胞加血管内皮生长因子C(VEGF-C)50 ng/mL诱导培养10 d,Western-blot法与免疫荧光染色法检测细胞中淋巴管内皮细胞标记物Prox-1和LYVE-1的表达。结果骨髓间充质干细胞呈现典型的梭形和旋涡状排列;经VEGF-C诱导后,细胞变短、呈多边形。流式细胞学显示,分离培养的骨髓间充质干细胞表面抗原CD29、CD90呈阳性表达,CD31、CD14呈阴性表达;Western-blot检测显示,经VEGF-C培养诱导后,细胞明显表达淋巴管内皮特异性标记物Prox-1和LYVE-1;免疫荧光染色显示,诱导后细胞均明显表达Prox-1和LYVE-1,而未分化的骨髓间充质干细胞Prox-1和LYVE-1均不表达。结论 VEGF-C可以诱导骨髓间充质干细胞表达淋巴内皮细胞特异性标记物,促进其向淋巴管内皮转化。

Objective To study the differentiation potentiality and inducement conditions of bone marrow mesenchymal stem cells（BMSCs）into lymphatic endothelial cells（LECs）,and to provide an ideal source of LECs for lymphatic regeneration and reconstruction.Methods BMSCs were isolated from the bone marrow of SD rats,and the surface antigens CD31,CD14,CD29 and CD90 on BMSCs were detected by flow cytometry.Purified BMSCs were cultured for 10 days in the conditioned medium with vascular endothelial growth factor C（VEGF-C）（50ng/mL）.Then,expressions of lymphatic endothelial cell markers,（prospero ho-meobox protein 1（Prox-1） and lymphatic vessel endothelial receptor 1（LYVE-1）,were detected by Western blot and immunofluorescence.Results BMSCs were typically fusiform in shape and circinate in arrangement,while they transformed into polygonal cells after co-culture with VEGF-C.Flow cytometry showed that expressions of surface antigens CD29 and CD90 on BMSCs were positive,however,CD31 and CD14 were negative.After induction with VEGF-C,Western blot and immunofluorescence showed that prox-1 and LYVE-1 were expressed in differentiated BMSCs,while they were not expressed in undifferentiated BMSCs in the control group.Conclusion BMSCs can be induced to express the lymphatic specific antigen and to differentiate into LECs by VEGF-C in vitro.