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MTT法广泛用于检测细胞存活和增殖。本文表明,在多种实验条件下,MTT法检测淋转的灵敏度都很低;而检测NK杀伤活性的灵敏度很高,但某些最佳实验条件与已有报道相异。1 实验方法1.1 MTT法检测淋巴细胞转化功能 取Balb/c小鼠脾脏经Ficoll分离后的淋巴细胞接种于96孔板。实验组加PHA,另设对照组。置37℃,5％CO_2中培养72或48h。培养结束前4h各孔加MTT液(5mg/ml)20μl。取出离心去上清,各孔加酸化异丙醇或DMSO溶解,酶标仪570nm处测OD值。转化功能以转化率表示:
MTT method is widely used to detect cell survival and proliferation. This article shows that under various experimental conditions, the sensitivity of MTT assay for detecting lymph node metastasis is very low. However, the sensitivity of detecting NK cytotoxic activity is high, but some of the best experimental conditions are different from those reported previously. 1 Experimental Methods 1.1 Detection of lymphocyte transformation by MTT assay The spleen of Balb / c mice was inoculated into 96-well plates with Ficoll-isolated lymphocytes. The experimental group plus PHA, another set of control group. Set 37 ℃, 5% CO_2 cultured 72 or 48h. 4h before the end of incubation plus MTT liquid (5mg / ml) 20μl. Remove the centrifuge to the supernatant, the wells were acidified with isopropanol or DMSO dissolved, microplate reader 570nm OD value. Conversion function expressed as conversion rate: