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Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium

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Summary

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-aminopterinthymidine (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.

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Authors and Affiliations

  1. Department of Cell Biology, The Research Institute for Tuberculosis and Cancer, Tohoku University, 4-1 Seiryomachi, 980, Sendai, Japan

    Noritsugu Yabe, Yutaka Matsuya & Isao Yamane

  2. Department of Bacteriology, Yokohama City University School of Medicine, 2-33 Urafune, Minamiku, 232, Yokohama, Japan

    Mitsuru Takada

Authors
  1. Noritsugu Yabe
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  2. Yutaka Matsuya
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  3. Isao Yamane
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  4. Mitsuru Takada
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Yabe, N., Matsuya, Y., Yamane, I. et al. Enhanced formation of mouse hybridomas without hat treatment in a serum-free medium. In Vitro Cell Dev Biol 22, 363–368 (1986). https://doi.org/10.1007/BF02623523

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  • DOI: https://doi.org/10.1007/BF02623523

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