Development of a serum-free medium for in vitro immune responses by using beta-cyclodextrin. Demonstration of the requirements for polyamines.
A serum-free medium has been developed which is able to support primary antibody responses by cultured murine lymphocytes. This medium is based on RPMI 1640 that is supplemented with beta-cyclodextrin, insulin, transferrin, albumin, low density lipoprotein, putrescine and L-alanine as substitutes for fetal calf serum. Omission of anyone of these components resulted in a marked decrease of antibody responses. By employing the serum-free culture conditions, it was clearly demonstrated that polyamines such as putrescine, spermidine and spermine had positive effects on the development of antibody-forming cells. This serum-free medium supported the antibody response to sheep erythrocytes, trinitrophenyl (TNP)-Ficoll or TNP-lipopolysaccharide as efficiently as 10% fetal calf serum-containing medium. In addition, murine lymphocytes proliferated in response to an antigen or a mitogen equally well in these two types of medium.
Department of Biotechnology, Faculty of Engineering, Okayama University, Japan.
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Ohmori
D000818:Animals; D000917:Antibody Formation; D000921:Antibody-Producing Cells / Q000166:cytology*; D003470:Culture Media; D003505:Cyclodextrins; D003912:Dextrins; D004912:Erythrocytes / Q000276:immunology; D005260:Female; D005936:Glucans / Q000494:pharmacology; D066298:In Vitro Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D011073:Polyamines / Q000494:pharmacology; D011700:Putrescine / Q000494:pharmacology; D013213:Starch; D013997:Time Factors; D047392:beta-Cyclodextrins
D003470:Culture Media; D003505:Cyclodextrins; D003912:Dextrins; D005936:Glucans; D011073:Polyamines; D047392:beta-Cyclodextrins; C031146:maltoheptaose; D013213:Starch; C031215:betadex; D011700:Putrescine