Formation of adherent monolayers of murine lymphocytes in vitro: the use of serum-free medium and concanavalin A-coated surfaces to promote adherence.
Murine thymus and spleen cells formed adherent monolayers in polystyrene tissue culture flasks when plated in serum-free medium. In the presence of 2% serum, thymus cells adhered poorly, but adherence was greatly enhanced if the flasks had been coated noncovalently with the lectin, concanavalin A. Adherence of leukemic lymphocytes (L1210) required both serum-free medium and concanavalin A-coated flasks; the extent of attachment was proportional to the concentration of the lectin used to coat the flasks at concentrations up to 0.1 mg/ml. Once L1210 cells had attached, they could not be removed by exposure to serum, ethylenediamine tetraacetic acid, trypsin, or alpha-methyl mannoside. Adherent L1210 cells remained capable of metabolism and proliferation during intervals of up to 7 days. The use of adherent monolayers for cytotoxicity assays was demonstrated by an assay for Pseudomonas aeruginosa toxin in EL4 murine leukemia cells.
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002478:Cells, Cultured; D003208:Concanavalin A / Q000494:pharmacology*; D003601:Cytotoxicity Tests, Immunologic; D007939:Leukemia L1210 / Q000503:physiopathology; D008214:Lymphocytes / Q000473:pathology / Q000502:physiology*; D051379:Mice; D013154:Spleen / Q000166:cytology; D013950:Thymus Gland / Q000166:cytology
D003208:Concanavalin A