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Establishment and Identification of HGPRT Deficient Hela Cell Line
Guo Huidong, Tergel, Shang Aiping, Zheng Wenjing, Liu Gang, Daorna, Wang Rongrong, Li Yao, Li Yu*
(Key Laboratory of Ministry of Education, Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University,Hohhot 010021, China
Abstract: To establish an HGPRT– deficient Hela cell line for creation of hybridoma to produce humanized
monoclonal antibody. Mutation of Hela cells was induced with MNNG and selected in gradient concentrations
of 6-TG. Then， the sensitivity of mutant cells to HAT medium was tested. After the stable Hela-HGPRT– cells were
developed， they were fused with human B lymphocytes and selected in HAT medium.Hela-HGPRT– cells could survive
in medium containing 20 μg/mL 6-TG in the long term and could not live in HAT medium. We also succeeded
in fusing human B lymphocytes with Hela-HGPRT– cells and the hybridoma cells could be continually cultured.
Using the methods of induction with MNNG and screening with 6-TG， we obtained a stable Hela-HGPRT– cell line
and it could be continually cultured. This Hela-HGPRT– cells could be used in the research of cell fusion.
monoclonal antibody. Mutation of Hela cells was induced with MNNG and selected in gradient concentrations
of 6-TG. Then， the sensitivity of mutant cells to HAT medium was tested. After the stable Hela-HGPRT– cells were
developed， they were fused with human B lymphocytes and selected in HAT medium.Hela-HGPRT– cells could survive
in medium containing 20 μg/mL 6-TG in the long term and could not live in HAT medium. We also succeeded
in fusing human B lymphocytes with Hela-HGPRT– cells and the hybridoma cells could be continually cultured.
Using the methods of induction with MNNG and screening with 6-TG， we obtained a stable Hela-HGPRT– cell line
and it could be continually cultured. This Hela-HGPRT– cells could be used in the research of cell fusion.
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