Comparison of Cul Tures of Human Lymphocytes Obtained Following Nh4Cl Induced Red Blood Cell Lysis and Ficoll-Hypaque Density Gradient Centrifugation

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Abstract
Lymphocyte thymidine uptake in leucocytes prepared by NH4Cl induced RBC lysis was compared with that of lymphocytes obtained following Ficoll-Hypaque centrifugation using polynomial regression dose response curves which adjusted for inherent subject variability. Eleven male subjects were studied. Heparinized blood was either layered on- Ficoll-Hypaque or treated with 0.87% NH4Cl to induce RBC lysis. Phytohemagglutinin-P (PHA) dose response curves were determined for each subject using triplicate 5 day cultures. For each subject, at each PHA concentration ranging from 12.5 to 167ug/ml, triplicate mean cpm of Ficoll-Hypaque treated lymphocytes and NH4C1 treated lymphocytes were compared by the 2 tailed Student's T test. Responses of cultures of NH4Cl treated lymphocytes were significantly different (2+ subject effect; s.e.=22,256; for NH.C1 treated lymphocytes, mean cpm=58,039+1022.8(PHA concentration)-5.32 (PHA concentration)2+ subject effect; s.e.=37,966. The p values from the F tests for significance of these quadratic regressions are 0.28, respectively. The s. e. of the quadratic regressions are different, F=2.91, d. f.=30,30; p4Cl treated lymphocytes1 dose response quadratic regression and its larger s. e. indicates that the NH4C1 treated lymphocyte culture dose response curve is less predictable than that of Ficoll-Hypaque treated lymphocytes.