摘要
本文用有限稀释法对ＷＥＨＩ１６４细胞进行了亚克隆，从１２株中筛选出一株对ＴＮＦ高度敏感的亚克隆细胞株ＷＥＨＩ－Ｃｌ。ＭＴｓ的还原产物具有良好的水溶性，ＭＴＳ／ＰＭＳ比色法与ＭＴＴ法相比较具有简便、快速的优点。我们首先将ＭＴＳ引入细胞毒的检测，用ＷＥＨＩ１６４－Ｃｌ亚克隆株建立了检测ＴＮＦ的ＭＴＳ／ＰＭＳ比色法。此方法不仅简便、快速．而且敏感度高（比常规Ｌ９２９细胞检测法敏感１０～１５倍）、特异性强（ＩＬ－１、ＩＬ－２、ＩＬ－６、ＰＨＡ、ＣｏｎＡ、ＬＰＳ对检测结果均无明显影响）。
By limiting dilution of WEHI164 murine fibrosarcoma cells.we bave obtained a cell line,WEHI164 sub-clone C1.which is extremely sensitive to human TNF. A new anaIog of MTT,MTS(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt)could be reduced by living cells to preducewater-soluble formazan in the presence of PMS(phenazine methasulfate),so MTS/PMS colorimetric assay is potential inbioassays. By introducing MTS/PMS into cytotoxicity assay,we have estasblished MTS/PMS colorimetric assay for TNFusing WEHI164 subelone C1 as target cells.The results demonstrated the methed was rapid. simple,sensitive(10 timesmore sensitive than actinomycin-D treated L929 bioassay for TNF),and specific(recombinant IL-la,IL-2.IL-6 andPHA-P, ConA had no significant effects on the WEHI-Cl assay).
出处
《免疫学杂志》
CAS
CSCD
北大核心
1994年第4期263-266,共4页
Immunological Journal
基金
上海市科技启明星计划(92QB01322)
关键词
比色法
细胞亚克隆
肿瘤坏死因子
Cytokine biOassay, Tumor necrosis factor,Cytotoxicity,MTS/PMS colorimetric assasy,WEHI 164 sub-clone
