骨髓CD_(34)^+细胞体外分化为血管内皮细胞的研究

Differentiation of bone marrow-derived CD-(34)^+ cells into endothelial cells in vitro

目的 研究骨髓CD3 4 + 细胞体外分化为内皮细胞的能力 ,并初步探讨其分化条件。方法 利用免疫磁珠法从骨髓中提取CD3 4 + 细胞 ,体外经血管内皮生长因子 (VEGF)、成纤维生长因子 (bFGF)及胰岛素样生长因子 1(IGF 1)等诱导分化后 ,观察细胞生长及形态变化 ,并通过检测细胞血管内皮生长因子受体 2 (VEGFR 2 )、Ⅷ因子相关抗原抗体表达和摄取乙酰化 低密度脂蛋白(Ac LDL)情况等对分化细胞进行鉴定。结果 用免疫磁珠法从骨髓中可以提取高纯度CD3 4 + 细胞 ,定向诱导后细胞呈卵石形 ,透射电镜显示细胞内具有特征性的W P小体 ,免疫荧光染色检测细胞相关抗原VEGFR 2、Ⅷ /vWF呈阳性 ,同时细胞摄取Ac LDL。结论 骨髓CD3 4 + 细胞是具有多向分化潜能的干细胞 ,经VEGF、bFGF、IGF 1等诱导 ,可以分化为血管内皮细胞 ,是理想的组织工程种子细胞。

Objective To study the differentiation of human bone marrow-derived CD 34+ cells into endothelial cells in vitro.Methods CD 34+ cells were isolated from human bone marrow by magnetic bead selection and induced in the presence of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF) and insulin-like growth factor-1 (IGF-1).The growth and morphology of CD 34+ cells were observed under a microscope.The differentiated cells were identified by measuring uptake of acetylated low density lipoprotein (Ac-LDL) and immunofluorescence staining for the expression of Ⅷ/vWF,vascular endothelial growth factor receptor-2 (VEGFR-2).Results CD 34+ cells could be purified by magnetic bead selection from human bone marrow.After being induced by VEGF,bFGF,IGF-1,the cells reformed clones with cobble-stone morphology and expressed characteristic Weibel-Paladle bodies under a transmission electronic microscope.The cells could be identified by uptake of Ac-LDL and the positive staining for VEGFR-2,Ⅷ/vWF.Conclusion Bone marrow-derived CD 34+ cells are one group of stem cells with the capacity of differentiating into multiple cells lineages.They can undergo differentiation into endothelial cells after induced by VEGF,bFGF,IGF-1,suggesting that bone marrow-derived CD 34+ cells might be a kinds of new seed cells of tissue engineering.