奶牛MTT比色法的最佳条件探讨

Research on Optimum Condition of MTT Assay in Milk Cow

为建立奶牛淋巴细胞增殖反应MTT比色法,对试验条件进行了研究。应用L16(45)正交试验,对影响MTT比色法的四个主要因素,包括ConA浓度、细胞浓度、培养液小牛血清浓度及培养时间进行了比较和探索。试验表明几个因素对其增殖反应都有显著影响(P<0.05),且最佳反应条件为:15μg/mL的ConA、1×106/mL细胞浓度、10%小牛血清及60h的培养时间;影响增殖反应的先后顺序为细胞浓度、ConA浓度、培养时间及血清浓度。

Four major factors which affected the peripheral blood lymphocyte proliferatation of milk cow including concentration of ConA?cell,lymphocyte and fatal calf serum had been studied.The test showed significantly different effect on lymphocyte proliferatation by MTT colormetric assay(P<0.05),and the best condition of reaction was 15μg/mL ConA,1×106/mL cell concentration,10%FCS and 60 hours,the order of four factors was cell concentration,ConA concentration,culture time and the FCS concentration.