论文部分内容阅读
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 investigate the potential and conditions of bone marrow mesenchymal stem cells differentiating into lymphatic endothelial cells and to provide an ideal source of lymphatic vessels for regeneration and reconstruction. Methods Bone marrow mesenchymal stem cells were isolated from SD rats. The surface antigens CD31, CD14, CD29 and CD90 of bone marrow mesenchymal stem cells were detected by flow cytometry. The purified bone marrow mesenchymal stem cells plus vascular endothelial growth factor C (VEGF-C) 50 ng / mL induction culture for 10 days, Western-blot and immunofluorescence staining of lymphatic endothelial cells markers Prox-1 and LYVE-1 expression. Results Bone marrow-derived mesenchymal stem cells showed typical spindle-shaped and swirling arrangement. After induced by VEGF-C, the cells became shorter and polygonal. Flow cytometry showed that the surface antigen CD29 and CD90 of the cultured BMSCs were positive and CD31 and CD14 were negative respectively. Western-blot results showed that the expression of CD29, Prox-1 and LYVE-1 were detected by immunofluorescence staining. Prox-1 and LYVE-1 were obviously expressed in all the cells after induction, but none of Prox-1 and LYVE-1 in undifferentiated BMSCs were not expressed. Conclusion VEGF-C can induce bone marrow mesenchymal stem cells to express lymphatic endothelial cell specific markers and promote their transformation into lymphatic endothelial cells.