Enhanced Formation of Mouse Hybridomas without HAT Treatment in a Serum-Free Medium

• Published in: In Vitro Cellular & Developmental Biology 1986-07, Vol.22 (7), p.363-368
• Main Authors: Noritsugu Yabe, Matsuya, Yutaka, Isao Yamane, Mitsuru Takada
• Format: Article
• Language: English
• Subjects: Aminopterin - pharmacology; Animal cells; Animals; B lymphocytes; Biological and medical sciences; Biotechnology; Cell cultures. Hybridization. Fusion; Cell Division; Cell fusion; Cell Fusion - drug effects; Cell growth; Cell hybridization, cell fusion, hybridomas; Cell Line; Cell lines; Culture Media; Cultured cells; Eukaryotic cell cultures; Feeder cells; Fundamental and applied biological sciences. Psychology; Humans; Hybridomas; Hybridomas - drug effects; Hypoxanthine; Hypoxanthines - pharmacology; Lymphocytes; Methods. Procedures. Technologies; Mice; Microelectromechanical systems; Molecular and cellular biology; Multiple Myeloma - metabolism; Spleen - cytology; Spleen cells; Thymidine - pharmacology
• ISSN: 0883-8364; 2327-431X; 1475-2689
• DOI: 10.1007/bf02623523

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.