Long-Term Culture of Glutamine Synthetase-Transfected HepG2 Cells in Circulatory Flow Bioreactor for Development of a Bioartificial Liver
| Autor: | Shin Enosawa, Tomoyuki Miyashita, Seiichi Suzuki, Xiao-Kang Li, Miyuki Tsunoda, Hiroshi Amemiya, Mitsugu Yamanaka, Shinya Hiramatsu, Naoko Tanimura, Takeshi Omasa, Kenichi Suga, Toshiharu Matsumura |
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| Jazyk: | angličtina |
| Rok vydání: | 2000 |
| Předmět: | |
| Zdroj: | Cell Transplantation, Vol 9 (2000) |
| Druh dokumentu: | article |
| ISSN: | 0963-6897 1555-3892 09636897 |
| DOI: | 10.1177/096368970000900520 |
| Popis: | Glutamine synthetase (GS) is involved in an accessory pathway of ammonia removal in mammals. To develop a bioartificial liver with a human cell line, GS gene was transfected into HepG2 cells, which had no ammonia removal activity. After culturing in the presence of methionine sulfoximine (MSX), a GS inhibitor, we obtained a MSX-resistant HepG2 subline (GS-HepG2), which had amplified GS gene; ammonia removal activity was estimated to be 1/7 of that of rat primary culture hepatocytes. The cells were cultured in a circulatory flow bioreactor for 109 days, while they multiplied from 5 × 10 7 to 4 × 10 9 cells. Three days after inoculation, the ammonia level of the culture medium was lowered to a level maintained thereafter, suggesting that using recombinant cell lines for bioartificial livers enables long-term repeated treatment for hepatic failure patient. Judging from the rate of decrease in the amount of the added ammonia, the ammonia removal capability of 4 × 10 9 GS-HepG2 cells was almost equivalent to 5 × 10 8 porcine hepatocytes inoculated into the circulatory flow bioreactor. Apart from their ammonia removal activity, GS-HepG2 cells eliminated human tumor necrosis factor-α (TNF-α). Cytokine removal therefore promises to be another useful property of bioreactor cells. |
| Databáze: | Directory of Open Access Journals |
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