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Induction of immunoglobulin-producing human peripheral blood lymphocytes in serum-free medium
Authors:Yasuo Tanno  Sumio Arai  Tamotsu Takishima
Affiliation:First Department of Internal Medicine, Tohoku University School of Medicine, Sendai 980, Japan
Abstract:Induction of immunoglobulin-secreting cells from human peripheral blood lymphocytes in a serum-free culture medium was studied. Albumin, transferrin, insulin and fibronectin can replace serum entirely for support of pokeweed mitogen (PWM)-stimulated B lymphocytes, measured by a reverse hemolytic plaque assay using protein A-coated red cells. In this serum-free system, growth and maturation to IgM and IgG secretion occur at the same or higher efficiency as in conventional serum-containing medium, with maximum numbers of plaque-forming cells on day 6 at optimal dose of PWM, 0.5 ~ 5 μg/ml. This system can be used to avoid the interference from undefined serum components.
Keywords:serum-free medium  plaque-forming cells  human peripheral lymphocytes  fibronectin  transferrin  pokeweed mitogen  PWM  pokeweed mitogen  Ig  immunoglobulin  DMEM  Dulbecco's modified Eagle's medium  M-DMEM  modified DMEM  BSA  bovine serum albumin  FCS  fetal calf serum  PFC  plaque-forming cells  HBSG  Hank's balanced salt solution containing 0.1% gelatin  Con A  concanavalin A  PHA  phytohemagglutinin
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