In vitro reactivity to concanavalin A of bovine lymphocytes after cryopreservation.
Cryopreservation of bovine peripheral lymphocytes and its effect on the in vitro response to concanavalin A tested in a microculture system is described. Using DMSO as cryoprotectant in the medium, the cells were cooled to -30 degrees C at 1.3 degrees C/minute and further to -80 degrees C at 6 degrees C/minute and then rapidly to -196 degrees C by dropping in liquid nitrogen. The cells were recovered by rapid thawing in water at 30-35 degrees C and washed twice before use in the stimulation test. Ten percent DMSO had a much better protective effect than 5%; addition of 25% fetal bovine serum to the freezing had no favourable effect. In most of the 16 animals used in the experiments the frozen lymphocytes gave the same or a higher response to Con A than those kept in the DMSO containing medium at 4 degrees C for two hours. The responses of the frozen cells were comparable to those of fresh lymphocytes (kept at 4 degrees C for two hours in medium without DMSO).
D000818:Animals; D002417:Cattle; D003208:Concanavalin A / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide; D005260:Female; D005615:Freezing; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000276:immunology*; D011309:Preservation, Biological
D003208:Concanavalin A; D004121:Dimethyl Sulfoxide