title: Application of a serum-free medium in the growth and differentiation of human peripheral blood lymphocytes.
abstract: To eliminate variability due to nonspecific stimulation or inhibition by different lots of fetal bovine serum, the activation of human peripheral blood mononuclear cells in a modification of Iscove's medium (medium C-IMDM) was compared with the routinely used Roswell Park Memorial Institute (RPMI)-1640 medium containing 15% fetal bovine serum. Stimulation of peripheral blood mononuclear cells was enhanced and occurred at lower mitogen concentrations in C-IMDM compared with cells grown in RPMI-1640 supplemented with fetal bovine serum. Maximum incorporation of [3H]thymidine following stimulation by concanavalin A, phytohemagglutinin, and pokeweed mitogen was more than twice the peak values obtained in RPMI-1640 supplemented with fetal bovine serum. Concentrations of concanavalin A and phytohemagglutinin required for maximum stimulation were 0.5 and 15 micrograms/ml, and 0.3 and 1.0 micrograms/ml . 5 X 10(5) cells in C-IMDM and RPMI-1640, respectively. Cells grown in C-IMDM responded to lower concentrations of pokeweed mitogen and optimal growth in the serum-free medium required 0.4 micrograms/ml . 5 X 10(5) cells. The stimulation of immunoglobulin-producing cells in C-IMDM was enhanced and occurred at lower concentrations of pokeweed mitogen. Less variability of growth (i.e., incorporation of [3H]thymidine) and immunoglobulin synthesis occurred in peripheral blood mononuclear cells cultured in different preparations of C-IMDM than that reported for cells cultured in RPMI-1640 supplemented with different lots or batches of fetal bovine serum. These data suggest that C-IMDM may be an alternative to media supplemented with fetal bovine serum.
mesh_terms: D000328:Adult; D002454:Cell Differentiation; D003208:Concanavalin A / Q000494:pharmacology; D003470:Culture Media; D005260:Female; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis; D008055:Lipids; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010835:Phytohemagglutinins / Q000494:pharmacology; D011043:Pokeweed Mitogens / Q000494:pharmacology; D012710:Serum Albumin, Bovine; D014168:Transferrin
chemical_list: D003470:Culture Media; D007136:Immunoglobulins; D008055:Lipids; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D014168:Transferrin; D003208:Concanavalin A; D012710:Serum Albumin, Bovine