title: Optimizing whole blood lymphocyte proliferation in the rat.
abstract: Use of whole blood for lymphocyte proliferation experiments have advantages over the usual isolated lymphocyte procedure. We have systematically optimized methodology for whole blood lymphocyte proliferation in male Sprague-Dawley rats. Both phytohemagglutinin (PHA) and concanavalin A (ConA) stimulate lymphocyte blastogenesis in whole blood. The response of rat lymphocytes to ConA at supra-optimum conditions doubled that of PHA in contrast to what is observed in human lymphocytes. Proliferation rates relate non-linearly to concentrations of both mitogens with profiles showing lag, log and decline phases with increasing concentrations. The effect of incubation period followed a similar pattern with an optimum growth rate at 72 h. Whole blood from adrenalectomized rats gave higher proliferation rates than blood from intact normal rats. Advantages of the technique include: use of as little as 8 microliters of blood for each determination; rats need not be killed, the technique is less cumbersome, less time consuming, and cheaper to carry out than the traditional isolated lymphocyte method.
authors.affiliation: Department of Pharmaceutics, State University of New York at Buffalo 14260, USA.
authors.forename: A A
authors.initials: AA
authors.lastname: Fasanmade
authors.forename: W J
authors.initials: WJ
authors.lastname: Jusko
mesh_terms: D000315:Adrenalectomy; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D003208:Concanavalin A; D007158:Immunologic Techniques; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D008297:Male; D010835:Phytohemagglutinins; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D010835:Phytohemagglutinins; D003208:Concanavalin A