Construction of hybrid liver support system using gene amplified recombinant mammalian cells

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In order to establish the cell line which was suitable for hybrid artificial liver support system, an ammonia metabolizing activity was given to Chinese Hamster Ovary (CHO) and HepG2 cell line using genetic engineering techniques. CHO-K1 and HepG2 cells were transformed with pSV2-GS vector which contains glutamine synthetase gene connected with SV40 or CMV promoter. The recombinant cell line was selected under the various concentrations of glutamine synthetase inhibitor, methionine sulfoximine (MSX). The obtained MSX tolerable cell line had the high glutamine synthetase activity and could remove the ammonium from medium. The kinetic parameters of this cell line were determined under 0,1, or 2mM ammonia concentration. The specific ammonium consumption rates of established CHO and HepG2 cell line during growth phase were about one fourth and one seventh of that of primary hepatocyte respectively.