title: New parental cell lines for generating human hybridomas.
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.
authors.affiliation: Department of Immunology, University of Kiel, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Hundhausen
authors.forename: R
authors.initials: R
authors.lastname: Laus
authors.forename: W
authors.initials: W
authors.lastname: Müller-Ruchholtz
mesh_terms: D002460:Cell Line; D003470:Culture Media; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007041:Hypoxanthine Phosphoribosyltransferase / Q000172:deficiency; D013937:Thymidine Kinase / Q000172:deficiency
chemical_list: D003470:Culture Media; D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase