Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium.
A newly developed, serum-free medium (NYSF-404) selects for antibody-producing hybridomas after fusion of antigen-sensitized mouse spleen cells with myeloma cell lines P3-X63-Ag8-U1 (P3-U1), P3-X63-Ag8-6.5.3 (Ag8.653), or P3-NSI/1-Ag4-1 (NS-1). Without the need for hypoxanthine-aminopterin-thymidine (HAT) selection of hybrid cells, frequency of hybridoma formation in medium NYSF-404 is higher (twice) than that in serum- and HAT-containing medium. Colonies developed upon limiting dilution in the presence of the mortal parent myeloma cells in medium NYSF-404 and pure culture of antibody-secreting cells could be subsequently established. The results suggest that fusions can be done in serum-free medium and that the clonal growth of hybridomas is dependent on factors produced by parent myeloma cells under serum-free culture conditions. Such factors seem deficient in serum- and HAT-containing medium or are masked by serum.
D000630:Aminopterin / Q000494:pharmacology*; D000818:Animals; D002455:Cell Division; D002459:Cell Fusion / Q000187:drug effects*; D002460:Cell Line; D003470:Culture Media; D006801:Humans; D006825:Hybridomas / Q000187:drug effects*; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology*; D051379:Mice; D009101:Multiple Myeloma / Q000378:metabolism; D013154:Spleen / Q000166:cytology; D013936:Thymidine / Q000494:pharmacology*
D003470:Culture Media; D007042:Hypoxanthines; D019271:Hypoxanthine; D000630:Aminopterin; D013936:Thymidine