MTS和CCK-8法检测番鸭胚胎成纤维细胞增殖的比较  

Comparison of Experimental Conditions between MTS and CCK-8 for Muscovy Duck Embryo Fibroblast Proliferation

作　　者：裴星朝 周禄强[1] 叶超[1] 李昂[1] 

机构地区：[1]福建农林大学动物科学学院,福建福州350002

出　　处：《中国家禽》2014年第11期14-17,共4页China Poultry

基　　金：国家水禽产业技术体系建设专项资金(CARS-43-5)

摘　　要：为探讨MTS法和CCK-8法检测番鸭胚胎成纤维细胞增殖的最佳试验条件,以12~13日胚龄番鸭鸭胚作为胚胎成纤维细胞原代培养的细胞来源,利用传至F2代的胚胎成纤维细胞为研究对象,对比MTS法和CCK-8法在不同孵育时间及不同检测波长条件下吸光光度值的差异,研究MTS法和CCK-8法检测番鸭胚胎成纤维细胞增殖活性的最佳检测时间、最佳检测波长,根据细胞数量与OD值的关系曲线确定两种试剂各自的最适细胞浓度。结果表明：MTS法的最佳波长为492 nm、CCK-8法为450 nm,MTS法的灵敏度稍高于CCK-8法;两种方法的最佳检测时间均为加入试剂后4 h;MTS法的最适细胞接种浓度为1.25×103~1×104个/mL,CCK-8法为2.5×103~1×104个/mL。MTS和CCK-8法均能较好地检测番鸭F2代胚胎成纤维细胞的体外增殖情况,相对于CCK-8法,MTS法更加经济,灵敏度更高。To determine optimal experimental conditions of MTS and CCK-8 methods for cell proliferation assays in muscovy duck embryo fibroblast,muscovy duck embryos of 12 to 13 days age were used as source of primary culture of embryo fibroblasts,and embryo fibroblasts of F2 generation were applied to compare the difference of OD value under different wavelengths between MTS and CCK-8,and determine the optimal detection time and measurement wavelength of muscovy duck embryo fibroblast proliferation activity with MTS and CCK-8,then confirm the optimal cell concentration of each method according to the curve between the number of cells and OD values. The results showed that the optimal measurement wavelength for MTS was 492 nm and for CCK-8 was 450 nm;and the sensitivity of MTS was slightly higher than that of CCK-8;The optimal detection time of two methods was 4 h;The appropriate cell inoculating concentrations were 1.25×103~1×104per milliliter and 2.5×103~1×104per milliliter for MTS and CCK-8,respectively. Both MTS and CCK-8 could accurately detect proliferation of F2 embryos fibroblast in vitro,but MTS was more economical and sensitive than CCK-8.

关 键 词：番鸭胚胎成纤维细胞 MTS CCK-8 细胞增殖 

分 类 号：S865.13[农业科学—野生动物驯养]