Comparison of cultures of human lymphocytes obtained following NH4Cl induced red blood cell lysis and Ficoll-Hypaque density gradient centrifugation

M. R. Simon, L. D. Roi, S. Desai, D. J. Salberg, N. R. Rose

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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 167 μg/ml, triplicate mean cpm of Ficoll-Hypaque treated lymphocytes and NH4Cl 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 NH4Cl treated lymphocytes, mean cpm=58,039 +1,002.8 (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, respecively. The s.e. of the quadratic regressions are different, F=2.91, d.f.=30,30; p4Cl treated lymphocytes' dose response quadratic regression and its larger s.e. indicates that the NH4Cl treated lymphocyte culture dose response curve is less predictable than that of Ficoll-Hypaque treated lymphocytes.

Original languageEnglish (US)
Pages (from-to)301-314
Number of pages14
JournalImmunological Communications
Volume12
Issue number3
StatePublished - 1983
Externally publishedYes

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Diatrizoate
Ficoll
Density Gradient Centrifugation
Erythrocytes
Lymphocytes
Centrifugation
Thymidine
Leukocytes
Students

ASJC Scopus subject areas

  • Immunology

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Comparison of cultures of human lymphocytes obtained following NH4Cl induced red blood cell lysis and Ficoll-Hypaque density gradient centrifugation. / Simon, M. R.; Roi, L. D.; Desai, S.; Salberg, D. J.; Rose, N. R.

In: Immunological Communications, Vol. 12, No. 3, 1983, p. 301-314.

Research output: Contribution to journalArticle

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T1 - Comparison of cultures of human lymphocytes obtained following NH4Cl induced red blood cell lysis and Ficoll-Hypaque density gradient centrifugation

AU - Simon, M. R.

AU - Roi, L. D.

AU - Desai, S.

AU - Salberg, D. J.

AU - Rose, N. R.

PY - 1983

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N2 - 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 167 μg/ml, triplicate mean cpm of Ficoll-Hypaque treated lymphocytes and NH4Cl 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 NH4Cl treated lymphocytes, mean cpm=58,039 +1,002.8 (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, respecively. The s.e. of the quadratic regressions are different, F=2.91, d.f.=30,30; p4Cl treated lymphocytes' dose response quadratic regression and its larger s.e. indicates that the NH4Cl treated lymphocyte culture dose response curve is less predictable than that of Ficoll-Hypaque treated lymphocytes.

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