Hep G2培养条件的摸索及其与CHL、3T3细胞代谢能力的比较  

Study on cultural condition of Hep G2 cells

作　　者：李欣[1] 熊习昆[1] 张文改[1] 王京滨[1] 陈秀娟[1] 

机构地区：[1]广东省疾病预防控制中心,广州510300

出　　处：《中国卫生检验杂志》2007年第8期1384-1386,1399,共4页Chinese Journal of Health Laboratory Technology

基　　金：广东省医学科学技术研究青年基金项目

摘　　要：目的:以MTT试验方法计算环磷酰胺(cyclophosphamide,CP)对Hep G2细胞和体外安全性评价常用细胞系CHL、3T3细胞的LC50,间接比较几种细胞系的代谢能力并确定Hep G2细胞适宜的培养试验条件。方法:用需代谢活化的阳性物CP以含血清和不含血清的培养液配制染毒液,按10.0、7.21、5.19、3.73、2.69、1.93、1.39、1.00 mg/ml浓度对CHL、3T3细胞和以不同培养液培养的Hep G2细胞染毒,在24 h时间点以MTT方法对细胞存活情况进行分析,计算LC50,比较各种细胞和不同培养条件下Hep G2细胞对CP的代谢活化水平,并观察不同培养条件下Hep G2细胞的生长状态。结果:MTT试验结果表明Hep G2细胞对CP的代谢能力最强,CHL细胞次之,而3T3细胞对CP的代谢能力最弱;3种不同培养液培养的Hep G2细胞无论染毒液是含血清的还是不含血清的均观察到CP对细胞的LC50:MEM>DMEM>RPMI 1640,Hep G2细胞在DMEM、RPMI 1640两种培养液中生长状态均良好,尤其是RPMI 1640培养的Hep G2细胞,在无血清的染毒液培养下既能获得较多的细胞又能表现出较强的代谢CP的能力。结论:Hep G2细胞对CP的代谢活化能力较CHL和3T3细胞强,以RPMI 1640培养的Hep G2细胞生长状态良好且较DMEM和MEM培养的细胞表现出更强的代谢活化CP的能力,能获得较理想的培养和试验结果。Objective：To calculate the LC50 of cyclophosphamide （CP） to Hep G2 cells and CHL 3T3 cells which are generally used in the assessment of safety, indirectly compare the metabolic activity among three cell lines, and determinate the ideally cultural and experimental condition of Hep G2 cells by MTT tests. Methods：CP,known to be metabolized to toxin, was prepared with MEM,DMEM and RPMI 1640 containing or not containing 10% newborn bovine serum respectively, administered to CHL, 3T3 cells and Hep G2 cells cultured with three different cell culture media at 10. 0,7.21,5.19,3.73,2. 69,1.93,1.39,1.00 mg/ml and the viability of cells was studied at 24 h with MTT test. Then LC50 was calculated. The metabolic activity was compared among three cell lines, and among three different cultured conditions of Hep G2 cells. The living state of Hep G2 cells in three different cell culture media was observed too. Results：MTT test： Hep G2 cells had the highest ability in metabolizing CP compared to the other two cells of CHL and 3T3, 3T3 cells were the lowest ones among them. The LC50 of Hep G2 cells to CP cultured with three kinds of culture media decreased from MEM to DMEM, then to RPMI 1640, weather the culture media contained newborn bovine serum or not. Hep G2 cells cultured in DMEM and RPMI 1640 all grew well, especially Hep G2 cells treated by CP prepared with RPMI 1640 containing no newborn bovine serum could get not only enough cells but also the ideal metabolic effecL Conclusion： The metabolic activity of Hep G2 cells to CP is higher than those of CHL,3T3 cells. Hep G2 cells cultured in DMEM and RPMI 1640 all grow well, especially Hep G2 cells cultured with RPMI 1640 showed stronger capacity in metabolizing CP than those cultured with MEM and DMEM. The results of culture and experiment are all satisfactory.

关 键 词：HEP G2细胞 CHL细胞 3T3细胞 环磷酰胺 

分 类 号：R155.3[医药卫生—营养与食品卫生学]