体外诱导骨髓CD34+细胞生成血管内皮细胞的方法

目的：体外分离狗骨髓CD34^ 细胞，建立诱导分化血管内皮细胞的方法，并进行生物学鉴定。方法：利用免疫磁珠法分离狗骨髓CD34^ 细胞，在体外经血管内皮细胞生长因子（VEGF）、内皮细胞生长因子（EGF）、碱性成纤维细胞生长因子（hFGF)诱导后，观察细胞生长状况，以光镜、电镜进行形态学鉴定，应用免疫组化方法检测冯维靳布兰德因子（Von Willebrand factor,vWF)表达。结果：光镜下细胞单层融合生长，呈铺路石样形态，单个核位子中央，细胞群体倍增时间为35h。电镜下可见到Weibel-Palade小体，在细胞胞浆vWF染色阳性。结论：采用免疫磁珠法可从骨髓获得高纯度的CD34^ 细胞，在体外诱导分化成血管内皮细胞。

Induction of CD34+ cell into endothelial cell in vitro

Objective To culture and observe the biological characteristics of endothelial cells derived from CD34 + cells isolated from dog bone marrow. Methods CD34 + cells were isolated from bone marrow by magnetic bead selection,and then was induced in the presence of vascular endothelial growth factor (VEGF), endothelial growth factor (EGF) and basic fibroblast growth factor(bFGF).The cells were identified by microscopy and transmission electron microscopy, and immunochemical staining with Von Willebrand factor (vWF) .Results The cells uniformly took on a cobblestone morphology undermicroscopy, the population double time of the cells was 35h.Undertransmission electron microscope, the cells displayed Weibel-Palade bodies. The cells are vWF positive. Conclusion It seems that magnetic bead selection is an effective ways to isolate CD34 + cells from bone marrow ,and CD34 + cells may be induced to form vascular endothelial cells.