摘要
目的:筛选出人脐静脉内皮细胞(HUVECs)原代、传代最适培养液.方法:0.02%II型胶原酶灌注脐静脉消化15min获得细胞,用不同的培养液培养.24h后观察细胞贴壁状况,细胞计数法得到贴壁细胞数量;内皮细胞标志性蛋白vWF进行细胞鉴定.传代培养后,用MTT法比较不同培养液对HUVECs活力的影响.结果:用含ECGS、20%FBS的ECM组进行原代培养,可获得贴壁细胞数量最多的HUVECs,且与其他组间有显著差异;用含30ng·mL-1 VEGF165、10%FBS的M199进行传代培养,HUVECs活力较ECM组无明显差异.结论:HUVECs原代、传代培养分别选用优化后的培养液,既保证原代HUVECs的质量和数量,又使传代培养成本降低60.8%.
Objective：Screen the optimum mediam in primary culture and subculture of HUVECs.Methods：Cells were obtained by 0.02%type II collagenase perfusion of umbilical vein to digest 15 min and cultured in different medium.Observe cell adherence status after 24 hand the number of cells was calculated by cell counting method;Identify endothelial cell by marker protein vWF.After subculture,the effect of different culture solution on the activity of HUVECs was compared via the MTT method.Results：The largest number of HUVECs were harvested using ECM medium with ECGS,20%FBS in primary culture,which exhibited significant differences with the other groups;HUVECs were subcultured by M199 medium containing30ng·mL-1 VEGF165 and 10%FBS,but the difference of HUVECs viability was not obvious compared with the ECM group.Conclusion：Use optimized medium for primary culture and subculture of HUVECs to ensure the quality and quantity of the original HUVECs and reduce the cost of passage culture by 60.8%.
作者
郭梦龙
包小丹
刘巧利
全克玲
井长勤
王文锋
GUO Menglong BAO Xiaodan LIU Qiaoli QUAN Keling JING Changqin WANG Wenfeng(College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, Chin)
出处
《河南师范大学学报（自然科学版）》
CAS
北大核心
2016年第5期112-116,共5页
Journal of Henan Normal University(Natural Science Edition)
基金
河南省科技厅科技攻关项目(142102310045
152102210126)
新乡医学院培育基金(20132D109)
