Effects of different protein supplements on mitogen responses of human peripheral blood lymphocytes |
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Authors: | Richard L. Rumley Stanley W. Chapman Marie L. Hoover Marvin A. Cuchens |
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Affiliation: | Veterans Administration Medical Center, and Departments of Medicine and Microbiology, University of Mississippi Medical Center, Jackson, MS 39216, U.S.A. |
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Abstract: | We evaluated the effects of 6 supplements often used in human lymphocyte cultures, including fetal calf serum, autologous human serum, pooled human AB serum, hypogammaglobulinemic human serum, bovine serum albumin and human serum albumin. Lymphocyte proliferation of unstimulated and mitogen activated peripheral blood mononuclear cells was measured by [3H]thymidine incorporation. The responses of cells stimulated with the T-cell mitogen phytohemagglutinin-P were significantly lower when cultured in bovine serum albumin supplemented media, but were otherwise not supplement dependent. In contrast, responses of cells stimulated with the B-cell mitogens Cowan I strain of S. aureus and antisera against the μ or δ chain of human immunoglobulin were significantly effected by supplement. Cultures containing fetal calf serum and bovine serum albumin had high background responses without a proportional rise in cellular proliferation when B-lymphocyte-specific mitogens were utilized. Autologous human serum and pooled human AB serum contained immunoglobulin which interacted with each of the B cell mitogens, thus limiting their usefulness as in vitro supplements. Cells grown in human serum albumin supplemented media had minimal background and high stimulated responses to B-cell mitogens. These results indicate that human serum albumin is an optimal supplement for in vitro human lymphocyte proliferative assays since it supports high stimulated cell responses with low background activity, is devoid of immunoglobulin and had minimal variability among lots. |
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Keywords: | peripheral blood mononuclear cells T lymphocytes B lymphocytes lymphocyte mitogens cellular proliferation serum-free media human serum albumin PBM peripheral blood mononuclear cells PHA phytohemagglutinin-P CISA HSA human serum albumin FCS fetal calf serum BSA bovine serum albumin ABS pooled AB human serum AHS autologous human serum HHS hypogammaglobulenemic human serum Ig immunoglobulin SI stimulation index cpm counts per minute Δ cpm background subtracted counts |
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