血管内皮细胞初始接种密度在细胞增殖和毒性检测中的重要性  

Effects of Initial Cell Seeding Density on the Evaluation of Vascular Endothelial Cell Proliferation and Toxicity

作　　者：霍雪萍[1] 张琳萍[2] 赵向绒[1] 刘勤社[3] 王海芳 HUO Xue-Ping;ZHANG Lin-Ping;ZHAO Xiang-Rong;LIU Qin-She;WANG Hai-Fang(Laboratory Center of Shaanxi Province People's Hospital,Xi’an 710068;Department of Integrated Traditional Chinese and Western Medicine,Medical School of Xi'an Jiaotong University,Xi'an 710061;Shaanxi Key Laboratory of Integrated Traditional and Western Medicine for Prevention and Treatment of Cardiovascular Diseases,Shaanxi University of Chinese Medicine,Xi'an 712046,China)

机构地区：[1]陕西省人民医院中心实验室,陕西西安710068 [2]西安交通大学医学部,陕西西安710061 [3]陕西省中西医结合心血管病防治重点实验室、陕西中医药大学,陕西西安712046

出　　处：《生物技术通讯》2018年第6期840-843,共4页Letters in Biotechnology

基　　金：国家自然科学基金(81573823);陕西省中医管理局平台项目(JCPT028)

摘　　要：目的:探讨不同接种密度对药物诱导血管内皮细胞增殖及损伤作用评价的影响,阐明优化细胞初始接种密度的重要性。方法:体外培养bEND.3小鼠血管内皮细胞,按照不同密度接种96孔培养板,给予药物处理,通过倒置显微镜观察细胞形态并拍照,采用WST-1比色法检测细胞活力,评价药物的促增殖作用或H_2O_2诱导氧化损伤的程度。结果:在进行72 h细胞增殖效应评价实验中,bEND.3细胞接种量应为3000～6000/孔,细胞密度过低、间隙较大或接种密度过大,均可能影响对药物效应的评价。在建立H_2O_2诱导细胞氧化损伤模型时,内皮细胞的接种量直接影响H_2O_2对细胞的损伤程度。结论:针对实验目的选择合适的细胞初始接种密度,对于提高实验结果的准确性和稳定性十分必要。Objective:To investigate the effect of cell seeding density on the evaluation of cell proliferation and cell injury.Methods:Murine brain microvascular endothelial(bEND.3)cells were cultured,and the cells in the logarithmic phase of growth were used in this experiment.The cell viability was determined using WST-1colorimetric method.The effect of initial cell seeding density on the evaluation of the pro-proliferation or toxic effects of drug substances was analyzed.Results:The seeding amount of bEND.3cells should be3000to6000/well in cell proliferation evaluation at72h after plating.The effect of drug substance on cell proliferation could be wrongly evaluated if the cell density was too low,or a very high seeding density was used.When H2O2induced oxidative damage model was established,the cell seeding density also directly affected the degree of H2O2damage to cells.Conclusion:Choosing a suitable initial cell seeding density based on the special experimental purpose is essential to improve the accuracy and stability of experimental results.

关 键 词：bEND.3细胞 细胞密度 细胞增殖 氧化损伤 

分 类 号：R-331[医药卫生] R-34