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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.