Improved MTT colorimetric assay for NKC activity
Zhang, P.; Zhang, C.; Liu, J.; Wei, D.; Zhao, Z.
Hua Xi Yi Ke Da Xue Xue Bao 27(2): 213-215
1996
ISSN/ISBN: 0257-7712 PMID: 9389047 Document Number: 457461
The established improved 3-(4, 5-dimethyl thiazoly) 2, 5-diphenyl-tetrazolium bromide (MTT) colorimetric assay for measuring the amount and function of living cell by the authors has the characteristics of simplicity, dependability and good repeatability within 3-12 hours. We have used this improved MTT colorimetric assay in measuring natural kill cell (NKC) activity of human peripheral blood. The results show that when the activities of effector cell(EC) and target cell (TC) are gtoreq 95%, and the density of EC is gtoreq 3 times 10-2/well and that of TC gtoreq 1.6 times 10-2/well, the number of living cell is positively correlated with the OD-570nm volume of purple formazan metabolite of MTT solution (r-e = 0.94, r-t = 0.97, P lt 0.001). When the ratio of EC/TC is 40:1, the NKC activity of peripheral blood of health adults is equal to that report by Zhang Chongjie et al. By this MTT colorimetric assay, the NKC activity of peripheral blood of 10 leukemic patients has been detected. The hivin x +- s of NKC activity is 34.86 +- 7.80%, which is positively correlated with the result (35.16 +- 8.4%) of the inhibition of 3H-TdR incorporation assay performed at the same time (r = 0.876, P lt 0. 001). MTT colorimetric assay for NKC activity avoids getting in touch with isotope. It is a convenient and valuable method.