MTT法检测鸡脾脏和外周血NK细胞杀伤活性的条件优化  

Optimal conditions in MTT assays to detect the cytotoxicity of NK cells in chickens

作　　者：李宏梅[1] 胡敬东[1] 郭慧君[1] 郑玉姝[1] 刘翠艳[1] 赵宏坤[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医学报》2009年第4期476-478,共3页Chinese Journal of Veterinary Science

基　　金：国家"十一五"科技支撑计划重大项目(2006BAK02A21);山东省科技重大专项(SDSP2005041005);山东农业大学青年科技创新基金项目(23478)

摘　　要：选取不同日龄的SPF鸡和肉鸡，用贴壁法从血液和脾脏分离出NK细胞作为效应细胞，选择瘤细胞MDCCMSB1和MDCC-MSBCU147分别作为靶细胞，二者按比例混合，孵育，利用MTT法检测靶细胞的活力反映效应细胞对靶细胞的杀伤活性，并对效靶比、作用时间进行比较。结果显示，MSB-1和CU147均可作为NK细胞良好的靶细胞，但CU147更稳定；效靶比为20：1-50：1，作用时间8-16h杀伤效果稳定；50％DMF20％SDS混合物作为裂解液能够对结晶彻底溶解，优于其他溶解试剂。The cytotoxicity of NK cells isolated from blood and spleen of different day-chickens as effect cells to target cells, MDCC-MSB1 and MSBCU142, were detected by MTT assays and evalue the ratio of effect cells and target cells and the time of incubation. The result indicated MSB1 and MSBCU147 could be better target cells in the assay of NK cells cytotoxicity and the optimal conditions were 20 ： 1-50 ： 1 as the ratio of effect cell and target cells and 8-16 h of incubation and the 20%SD-50%DMF mixture as lytsis to MTT product. This will be help to analyze the cytotoxicity of NK cells in chickens.

关 键 词：鸡 MTT法 NK细胞杀伤活性 

分 类 号：S831[农业科学—畜牧学] S852.2[农业科学—畜牧兽医]