人脐静脉内皮细胞体外分离培养的改进及其鉴定

Improved Cultivation of HUVECs in Vitro and Their Identification

[目的]优化建立人脐静脉内皮细胞(HUVEC)体外原代及传代培养的方法,并对培养的细胞进行鉴定,为组织工程血管提供种子细胞.[方法]改进的胶原酶I型灌注法分离HUVEC,用含胎牛血清及内皮细胞生长因子的RPMI-1640培养基培养,光镜、免疫组化法进行生物学鉴定.[结果]分离的HUVEC体外培养7～10 d左右可长成单层,光镜下为多角形,呈铺路石状排列,人第Ⅷ因子相关抗原免疫组化检测阳性.[结论]改进的胶原酶I型灌注法所分离细胞数量多,易成活,且为具有细胞生物学功能的HUVEC.

ObjectiveTo establish a better method for cultivating primary and subcultured HUVECs in vitro and identify the cultural cells, so as to provide seed cells for tissue engineering vascular grafts.Improved collagenase typeⅠ perfusion method was used to isolate HUVECs , then cultivated in RPMI-1640 containing fetal bovine serum and endothelial cell growth factors, identified the biology of HUVEC with the light microscope and immunohistochemical method.Isolated HUVECs grew to monolayer after 7~10 days by cultivation in vitro. They presented polygonal and arranged like pavement stones under the light microscope. Correlation antigens of human Ⅷ factor were positive.[Conclusion]Cells isolated by improved collagenase typeⅠ perfusion method was more and easy to survive .They was proved to be HUVECs and present relative biological functions.