EA.HY926人脐静脉内皮细胞株与原代细胞生物特性的比较研究

Comparative study of biological characteristics between primary human umbilical vein endothelial cells and human umbilical vein endothelial cell line

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, disadvantages and the biological characteristics on the basis of the cultivation of EA. HY926 human umbilical vein endothelial cell lines and primary cell line. Methods Compared two umbilical vein endothelial cell＇s differences in morphology, growth curve, etc by means of light and electronic microscope morphological identification, VIII factor related antigen immunohistochemical examination and the determination of growth curve. Results Under transmission electron mi croscopy,more cross-section and longitudinal section of webel-palade bodies （WPB） could be found in primary HUVEC sample than in EA.HY926 cell lines. The average optical density value of EA. HY926 human umbilical vein endothelial cell line and the original generation of HUVECs was respectively （2 234.02 ±78.78） and （4 138.8±594.01） by the analysis of Immunohistochemical staining Image-Pro Plus 6.0 soft-ware. The immunohistochemical staining of the original generation of HUVEC was better than EA. High HY926 cell lines, with significant difference （P 〈0.001）. The growth curve of EA. HY926 human umbilical vein endothelial cell line was more stable than the original generation HUVECs. Conclusion Primary cells cultivation was time-consuming and expensive, but with the characteristics of short growth period, mixed cells pollution and limited cell proliferation ability. EA. HY926 cell lines are not porfound in the physiological characteristics, but with simple culture method and high survival rate,and it＇s a good source for basic research related to tissue engineering and cellular medicine.