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      New parental cell lines for generating human hybridomas

      , ,
      Journal of Immunological Methods
      Elsevier BV

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          Abstract

          In contrast to the success achieved with the production of hybridomas in the mouse system, creating human hybridomas is problematic. The reason is believed to be a lack of suitable malignant human cell lines. The work presented here demonstrates the establishment of three human parent cell lines--two of which are of T cell origin--by installing hypoxanthine guanosine phosphoribosyl transferase (HGPRT) and/or thimidine kinase (TK) deficiencies into the leukemic cell lines REH, 1301 and SKW-3. In order to isolate true hybridomas, selection procedures must guarantee complete death of the enzyme-deficient malignant parent cells. In this respect sublines with a combined HGPRT and TK deficiency proved to be superior to those with only one enzyme deficiency, especially in combination with the newly developed hypoxanthine/aminopterine/thymidine/azaserine (HATA) selection medium. However, selection media should be of low nonspecific toxicity. This was shown to be a particular property of the thymidine-free azaserine/hypoxanthine (AH) selection medium. Preliminary data show an extraordinary ability of one subclone of the hypoxanthine guanosine phosphoribosyl transferase-negative T cell line SKW-3 to generate human T-T hybridomas. They are of a stable, nearly tetraploid karyotype and express new surface antigens, thus providing new possibilities for the investigation of human T lymphocyte function by means of hybridoma technology.

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          Author and article information

          Journal
          Journal of Immunological Methods
          Journal of Immunological Methods
          Elsevier BV
          00221759
          August 1992
          August 1992
          : 153
          : 1-2
          : 21-29
          Article
          10.1016/0022-1759(92)90301-9
          1517591
          08445fab-8a01-4563-adc3-ce1697af0375
          © 1992

          https://www.elsevier.com/tdm/userlicense/1.0/

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