Establishment of a Human TGF-β1 Producing Cell Line by Gene Transfer

초록

Transforming Growth Factor-β1 (TGF-β1) is a pleiotrophic polypeptide involved in various biological activities including cell growth, differentiation and extracellular matrix deposition. Since TGF-β1 plays important roles in wound healing processes as well as in immune regulation, TGF-β1 is considered as a potential therapeutic agent for wound healing or organ transplantation. To obtain large quantity of human recombinant TGF-β1, an expression plasmid was constructed and transfected into Chinese hamster ovary (CHO) cells, A TGF-β1 producing clone was selected by measuring TGF-β1 levels in the medium and incubated in increasing concentrations of methotrexate up to 60 μM for the amplification of TGF-β1 cDNA in conjunction with a dihydrofolate reductase (dhfr) gene. The cell line selected in the medium containing 60 μM methotrexate produced TGF-β1 at the concentration of 12 ng/ml per 105 cells/day, which is approximately 9 fold over that of the transfected cells incubated without methotrexate. The selected cell line showed a slow growth rate and changed morphology. Overall, the established CHO cell line in this study would be useful to produce recombinant human TGF-β1.

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