大鼠脐血来源单个核细胞诱导为内皮祖细胞的免疫荧光检测

目的脐血（umbilical cord blood，UCB）中含有大量内皮祖细胞（endothelial progenitor cells，EPCs），内皮祖细胞（EPCs）是血管内皮细胞的前体细胞，有分化为血管内皮细胞的能力，为骨组织工程血管化带来新的途径。本实验研究大鼠脐血来源的单个核细胞体外诱导为EPCs的可能性。方法分离大鼠脐血单个核细胞，血管内皮诱导液（添加10％胎牛血清，1％青霉素，1％链霉素，VEGF20ug／L，IGF-12ug／L，bFGF2ug／L，EGF20ug／L）培养分化，于3周、6周进行免疫荧光染色检测贴壁细胞CD34、CD133、Flk-1、vWF表面标志。结果培养三周单个核细胞CD34、CD133、Flk-1、vWF抗体免疫荧光检测均为阳性；六周免疫荧光染色可见CD34、Flk-1、VWF组为阳性，CD133组部分为阳性，阴性对照组未发现阳性细胞。结论大鼠脐血来源的单个核细胞在体外诱导3周可以分化为EPCs，6周部分细胞分化为成熟血管内皮细胞。

Immunofluorescent Staining on Differentiation of Mononuclear Cells from Rat Umbilical Cord Blood into Endothelial Progenitor Cells

Objective Umbilical cord blood（UCB）is best containing a lot of endothelial progenitor cells（EPCs）, they are progenitors of vascular endothelial cells （VECs）and capable of differentiation into VECs. It will lead to a new strategy to promote angiogenesis in bone tissue engineering. The purpose of this study was to analyze the potentiality on differentiation of mononuclear cells from rat umbilical cord blood into endothelial progenitor cells in vitro. Methods To isolate mononuclear cells from umbilical cord blood, they were cultured in conditioned medium（supplemented with 10% FBS, 1% penicilin, 1% phytomycin, VEGF 20 u g/L, IGF-1 2 u g/L, bFGF 2 u g/L, EGF 20 u g/L）. The immunofluorescent staining were used to observe expressions and distributions of some special surface marker, such as CD34, CD133, Flk-1 and vWF on adherent cells after 3 weeks and 6 weeks. Result The results of the immunofluorescent staining were positive on CD34, CD133, Flk-1 and vWF after 3 weeks, but their results were only positive on CD34, Flk-1 and vWF after 6 weeks, the part of adherent cells were positive on CD133, the positive cells were not found in the control group. Conclusion The mononuclear cells from rat umbilical cord blood were induced into endothelial progenitor cells after 3 weeks in vitro.the part of adherent cells differentiated into mature vascular endothelial cells.