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目的比较培养EA.HY926人脐静脉内皮细胞株与原代脐静脉内皮细胞(HUVECs)的优缺点及生物学特性。方法通过倒置显微镜、透射电镜形态学鉴定、Ⅷ因子相关抗原免疫组织化学检查及生长曲线测定,比较2种脐静脉内皮细胞的形态、生长曲线等差异。结果透射电镜观察到原代HUVECs中发现较多WebelPalade小体,而EA.HY926细胞株较少见。免疫组织化学染色Image-Pro Plus 6.0软件进行分析EA.HY926人脐静脉内皮细胞株与原代HUVECs平均光密度值分别为(2 234.02±78.78)、(4 138.80±594.01)。原代HUVECs的染色程度明显较EA.HY926细胞株高,差异有统计学意义(P<0.01)。EA.HY926人脐静脉内皮细胞株生长曲线较原代HUVECs稳定。结论培养原代HUVECs耗时长,生长期较短,易出现杂细胞的污染,细胞的增殖能力有限,花费昂贵。EA.HY926细胞株虽生物特性较差,但其操作简便,具有一定程度的原代HUVECs所具有的生物特性,细胞均一以及增长旺盛,可用于较复杂、细胞创伤较大的体外内皮细胞的研究。
Objective To compare the advantages and disadvantages and biological characteristics of EA.HY926 human umbilical vein endothelial cell line with primary human umbilical vein endothelial cells (HUVECs). Methods The morphology, growth curve and other differences of two kinds of umbilical vein endothelial cells were compared by inverted microscope, transmission electron microscopy morphological identification, immunohistochemical examination of Ⅷ factor related antigen and growth curve. Results Transmission electron microscopy revealed more WebelPalade bodies in primary HUVECs, while EA.HY926 cells were rare. The average optical density of EA-HY926 human umbilical vein endothelial cell line and primary HUVECs were (2 234.02 ± 78.78) and (4 138.80 ± 594.01), respectively, by immunohistochemistry Image-Pro Plus 6.0 software. The staining of primary HUVECs was significantly higher than that of EA.HY926 cells (P <0.01). The growth curve of EA.HY926 human umbilical vein endothelial cell line was more stable than that of primary HUVECs. Conclusion The culture of primary HUVECs takes a long time and has a short growth period. It is easy to appear the contamination of the extracellular cells. The proliferation of cells is limited and costly. Although the biological characteristics of EA.HY926 cell line is poor, its operation is simple, and it has a certain degree of biological characteristics, uniform cells and strong growth of primary HUVECs and can be used for the study of more complex and traumatic endothelial cells in vitro. .