[3H-thymidine-paper disc-counting of in vitro lymphocyte transformation demonstrated on phytomitogen-activated rabbit peripheral blood lymphocytes (author's transl)].
A simple routine method for the phytomitogen-(PHA-, Con A-) induced stimulation of purified rabbit peripheral blood lymphocytes is described. The lymphocytes are cultivated in closed polypropylene Eppendorf tubes at 37 degrees C, the mitogen answer is quantified by determination of 3H-thymidine (3HT) incorporation by means of liquid scintillation spectrometry of cell-sediment layered filter paper discs. The optimal experimental conditions for each step of the test procedure are given. The mean variation coefficients of triplicate determinations of non-stimulated negative controls (n = 28) and mitogen-activated lymphocyte cultures (n = 56) are 17.0 (6-28) and 5.6 (1-13)%, respectively.
D000818:Animals; D002469:Cell Separation; D003208:Concanavalin A / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D011817:Rabbits / Q000276:immunology; D013936:Thymidine / Q000378:metabolism*; D014316:Tritium
D010835:Phytohemagglutinins; D014316:Tritium; D003208:Concanavalin A; D004247:DNA; D013936:Thymidine
Anwendung der 3H-Thymidin-Papierplättchen-Modifikation des Lymphozytentransformationstestes am Beispiel Phytomitogen-aktivierter peripherer Kaninchenblutlymphozyten.