体外诱导骨髓间充质干细胞分化为血管内皮细胞的实验研究

The Empirical Study of Differentiation of Bone Marrow Mesenchymal Stem Cells into Cells of Endothelial Lineage in vitro

目的探讨体外诱导骨髓间充质干细胞分化为血管内皮细胞的可行性。方法分离培养骨髓间充质干细胞,鉴定其表面抗原,用含2%胎牛血清以及50 ng/m l血管内皮生长因子的条件培养基诱导,诱导后细胞通过内皮细胞标志物KDR、FLT-1以及v-WF染色鉴定。结果骨髓间充质干细胞表面抗原CD29、CD44、CD105阳性,CD34、CD45、CD106以及HLA-DR阴性。诱导后细胞形态明显改变,内皮细胞标志物KDR、FLT-1以及v-WF染色结果阳性。结论骨髓间充质干细胞在体外具有分化为血管内皮细胞的能力,可以作为干细胞治疗的一种有效来源。

Objective To investigate the in vitro differentiation of bone marrow mesenchymal stem ceils into cells of vascular endothelial lineage. Methods Ceils were isolated from bone marrow, then were characterized by flow cytometry using a panel of monoclonal antibodies. Differentiation into endothelial-like cells was induced by cultivation of confluent cells in the presence of 2% fetal calf serum and 50 ng/ml vascular endothelial growth factor. These cells were characterized by immunohistochemistry stain. Results Isolated MSCs were positive for the markers CD105,CD29 and CD44 and negative for typical hematopoietic and endothelial markersCD34,CD45,CD106 and HLA-DR. The morphology of induced ceils was obviously changed. Immunohistochemistry stain analysis of the confluent cells in situ showed positive for endothelial-specific markers like KDR and FLT-1, and immunofluorescence analysis showed typical expression of the von Willebrand factor. Conclusion The bone marrow mesenchymal stem cells have the capability to differentiate ceils of endothelial lineage in vitro. The bone marrow MSCs may be a useful source of stem cell therapy.