China Animal Husbandry & Veterinary Medicine ›› 2011, Vol. 38 ›› Issue (12): 92-95.

• 生物技术 • Previous Articles     Next Articles

Establishment of Hypoxanthine Guanine Phosphoribosyl Transferase Mutant from Human Liver Cancer Cell Line HepG2

Tergel, TAI Da-peng, LIU Gang, Daorna, WANG Rong-rong, GUO Hui-dong, LI Yao, LI Yu   

  1. Key Laboratory of Ministry of Education,Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University,Hohhot 010021,China
  • Received:1900-01-01 Revised:2011-06-07 Online:2011-12-20 Published:2011-12-20

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

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