摘要
以牛皱胃平滑肌细胞为研究对象,采用MTT法和CCK-8法检测添加不同浓度β-羟丁酸(β-hydroxybutyricacid,β-HB)对牛皱胃平滑肌细胞活性的影响。通过接种不同的细胞密度((0.2、0.5、1、2、3)×105个/mL),加入MTT/CCK-8孵育后,在不同的波长下读取D值并进行分析。确定CCK-8法的最佳检测波长为450nm,最佳细胞浓度为1×105个/mL,最佳检测时间为4h;MTT法的最佳检测波长为490nm,最佳细胞浓度为3×105个/mL。在检测β-羟丁酸对牛皱胃平滑肌细胞活性的影响时,2种方法的结果对比发现CCK-8法的数据偏差小,准确性高,灵敏,优于MTT法。
To investigate the optimal experimental conditions of MTT and CCK-8. Different con- centration abomasum smooth muscle cells（（0.2,0.5,1,2,3） ×10^5/mL） were incubated with dif- ferent concentrations of β-hydroxybutyric acid（β-HB）, and the optimal absorbance wavelenght was measured by CCK-8 or MTT to determine the abomasum smooth muscle cell viability of dairy cow. We found that the optimal absorbance wavelenght was measured at 490 nm and the optimal concentration of cell was 3 ×10^5/mL in the MTT assay,and the optimal absorbance wavelenght was measured at 450 nm,the optimal concentration of cell was 1 ×10^5/mL and the optimal reac- ting time was 4 hours in the CCK-8 assay. These results suggest that CCK-8 assay leads to a smaller deviation,the accuracy and sensitivity is better than MTT assay for determining cell via- bility of abomasum smooth muscle cell pretreated with different concentrations of β-hydroxybu- tyric acid in vitro.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2013年第3期436-438,449,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31101865)
