Lymphoblastic transformation assay of sheep peripheral blood lymphocytes: a new rapid and easy-to-read technique.
A new micro-method was used to evaluate in vitro sensitivity of ovine peripheral blood lymphocytes (PBL) to different non specific mitogens (pHA, Con A, PWM) and to investigate the interest of a colorimetric assay for measurement of transformed lymphocytes. The results showed that sheep PBL in flat-bottomed microplates responded optimally at a cell density of 8 X 10(6) cells/ml to PHA (2.5 micrograms/ml), Con A (5 micrograms/ml) and PWM (5 micrograms/ml). The colorimetric assay using a tetrazolium salt (MTT), for measuring the transformed lymphocytes, is very well correlated with the classical method of [3H]thymidine incorporation. This new revelation technique of the mitogenic response improve the technical value of the assay, which is more rapid and easy-to-read, without diminishing the biological value.
D000818:Animals; D002455:Cell Division; D002470:Cell Survival; D003124:Colorimetry; D003208:Concanavalin A / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis; D005260:Female; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000378:metabolism*; D008215:Lymphocytes, Null; D008297:Male; D008934:Mitogens / Q000494:pharmacology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology; D012756:Sheep; D013778:Tetrazolium Salts; D013844:Thiazoles; D013936:Thymidine / Q000378:metabolism
D008934:Mitogens; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D013778:Tetrazolium Salts; D013844:Thiazoles; D003208:Concanavalin A; D004247:DNA; C022616:thiazolyl blue; D013936:Thymidine