Abstract: To establish a hypoxanthine guanine phosphoribosyl transferase(HGPRT) deficient cell line from HepG2 cell line,so as to provide parent cells for hybridoma and monoclonal antibody study. The HepG2 cells were cultured with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) for mutation and selected by increasing concent ration of 6-thioguanine (6-TG). The 6-TG resistant mutant cells were cultured in the medium containing 6-TG or HAT. After four months of screening,a deficient cell line was established. It could grow vigorously in the medium containing 20 μg/mL 6-TG,while it all died within 12 days in the medium containing HAT. Karyotypic analysis showed that the number of the chromosomes of the deficient cells is between 32 and 88, with the modal number being 48. The 6-TG resistant and aminopterin-sensitive character of the mutant cell line reveals that it is deficient in HGPRT. The cell line established in this study will be a good parent cell line for preparation of hybridoma in the future.

Key words: cell mutagenesis; HepG2; HGPRT-defective cell line