应用MTT测定PBMC增殖最佳条件的研究

The Study of the Optimum Condition to Measure the Proliferation of PBMC by MTT

对ＭＴＴ（噻唑蓝）测定离休ＰＢＭＣ（外周血单核细胞）增殖最佳实验条件进行了研究，结果表明，在测定由不同有丝分裂原（ＰＷＭ、ＳＥＢ和ＣｏｎＡ）诱导的ＰＢＭＣ增殖中，ＭＴＴ的Ｏ．Ｄ值和表示３￣Ｈ一ＴｄＲ摄取量的ｃｐｍ值呈正相关（ｒ＝０．９１５～０．９８２，ｐ＜０．０５，ｐ＜０．０１）。在ＰＢＭＣ密度为２．５×１０￣５／孔、孵育时间为４ｄ～６ｄ的条件下Ｐ／Ｈ比值最大，此外，细胞培养基中的血清种类及其浓度也是重要的影响因素。ＭＴＴ的浓度及催化形成噻唑盐的时间在达４ｈ后对Ｐ／Ｎ比值影响不大。所有这些表明，只要选择最佳条件，应用非同住素标记的ＭＴＴ法可在科研和常规检验时测定畜禽ＰＢＭＣ增殖。

This page reports the study of the optimum condition for the measurementof PBMC proliferation by MTT assay,a colorimetric methd.In a dynamic measure of theproliferation of PBMC induced by PWM,SEB and ConA,there was a strong regression be-tween the O.D in MTT and the cpm in 3H一TdR uptake(r=0. 915～0. 982,p<0.05and p<0.01).The greatst value of P/N under the condition of the cell一density in 2×105/well,the cell culture for 4～6 days and MTT一culture for 4～6 hours was observed.In ad-dition,the KInds and concentration of serums in the culture medium,but not concentrationof MTT in the medium,showed a significant influence on the value of P/N. All these resultsindicated that under the optimum condition the MTT assay could be an easy and inexpensivemethod to measure the proliferation not only of cell lines but also of PBMC in the study ofcellular immunoloy of livestock and pouitry.