| RRC ID |
68483
|
| 著者 |
Enosawa S, Miyashita T, Suzuki S, Li XK, Tsunoda M, Amemiya H, Yamanaka M, Hiramatsu S, Tanimura N, Omasa T, Suga K, Matsumura T.
|
| タイトル |
Long-term culture of glutamine synthetase-transfected HepG2 cells in circulatory flow bioreactor for development of a bioartificial liver.
|
| ジャーナル |
Cell Transplant
|
| Abstract |
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 x 10(7) to 4 x 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 x 10(9) GS-HepG2 cells was almost equivalent to 5 x 10(8) porcine hepatocytes inoculated into the circulatory flow bioreactor. Apart from their ammonia removal activity, GS-HepG2 cells eliminated human tumor necrosis factor-alpha (TNF-alpha). Cytokine removal therefore promises to be another useful property of bioreactor cells.
|
| 巻・号 |
9(5)
|
| ページ |
711-5
|
| 公開日 |
2000-1-1
|
| DOI |
10.1177/096368970000900520
|
| PMID |
11144971
|
| MeSH |
Air Movements
Ammonia / metabolism
Bioreactors*
Cell Culture Techniques / methods*
Cell Line
Glucose / metabolism
Glutamate-Ammonia Ligase / genetics*
Glutamate-Ammonia Ligase / metabolism
Humans
Liver, Artificial*
Recombinant Proteins / metabolism
Transfection*
Tumor Necrosis Factor-alpha / metabolism
|
| IF |
3.341
|
| リソース情報 |
| ヒト・動物細胞 |
Hep G2(RCB0459) |
