Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2016, Vol. 32 ›› Issue (26): 21-26.doi: 10.11924/j.issn.1000-6850.casb16060080

Special Issue: 生物技术 畜牧兽医

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Application of MTT Method for Detecting Rat Peripheral Blood Lymphocyte Proliferation

Liang Hongwei1, Zhu Wenyu1, Feng Bo1,2, Wang Jianfang1, Zhang Tao1, Mu Xiang1   

  1. (1Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Beijing University of Agriculture, Beijing 102206;2College of Veterinary Medicine, China Agricultural University, Beijing 100193)
  • Received:2016-06-16 Revised:2016-09-11 Accepted:2016-08-30 Online:2016-09-21 Published:2016-09-21

Abstract: In order to evaluate cellular immune state more accurately and simply, we used MTT colorimetric analysis to detect the proliferation of peripheral blood lymphocytes in rats, and analyzed the optimum conditions of the experiment. Then we applied MTT method to the Transwell co-culture system in order to assess the effect of infecting endothelial cells by H9N2 AIV on the proliferation of lymphocytes which crossing endothelial cells. Results showed that the optimum culture time was 48 h when inoculating 1.25×106 lymphocytes per milliliter into the RPMI 1640 medium with 10% FBS in a 96 hole cell culture plate. The proliferation of lymphocytes was inhibited apparently after infected by virus. Lymphocytes and virus co-incubation reduced the amount of live virus with intact endothelium. On the contrary, this phenomenon disappeared. The result indicated that as one of the evaluation indexes of cellular immunity, lymphocytes proliferation could be detected by the optimized MTT method.