Kino Y, Sawa M, Kasai S, Nakazawa E, Kato K, Yamamoto T, Mito M
Second Department of Surgery, Asahikawa Medical College, Hokkaido, Japan.
Cell Transplant. 1996 Sep-Oct;5(5 Suppl 1):S35-7. doi: 10.1016/0963-6897(96)00089-9.
We have investigated the availability of multiporous microcarriers (MCs) for immobilizing isolated rat hepatocytes, but the pore size of MCs was too small (35 microns) for hepatocyte immobilization. In this study, we immobilized isolated rat hepatocytes on MCs with larger pores, and evaluated their metabolic activity. Isolated hepatocytes were immobilized on MCs precoated with collagen by the intermittent stirring method and by aspiration, and the cell-protein content per 100 mg MCs was determined for comparison of these methods. Metabolic activity was evaluated by analyzing NH3 metabolism, urea nitrogen synthesis and glucose synthesis. The aspiration method immobilized significantly more of hepatocytes on MCs than the intermittent stirring method (p < 0.05). A stationary culture of hepatocytes immobilized on MCs showed a similar NH3 metabolism to monolayer cultured hepatocytes, and hepatocytes immobilized on MCs in a floating culture showed significantly higher NH3 metabolism than those in a stationary culture (p < 0.01). However, monolayer cultured hepatocytes showed higher glucose synthesis than hepatocytes immobilized on MCs in a stationary culture (p < 0.01). In conclusion, hepatocytes immobilized on MCs proved to be useful as a bioreactor in a hybrid artificial liver.
我们研究了用于固定分离的大鼠肝细胞的多孔微载体（MCs）的可用性，但MCs的孔径对于肝细胞固定来说太小（35微米）。在本研究中，我们将分离的大鼠肝细胞固定在具有更大孔径的MCs上，并评估它们的代谢活性。通过间歇搅拌法和抽吸法将分离的肝细胞固定在预涂有胶原蛋白的MCs上，并测定每100mg MCs的细胞蛋白含量以比较这些方法。通过分析NH3代谢、尿素氮合成和葡萄糖合成来评估代谢活性。抽吸法比间歇搅拌法在MCs上固定的肝细胞显著更多（p<0.05）。固定在MCs上的肝细胞的静态培养显示出与单层培养的肝细胞相似的NH3代谢，并且在悬浮培养中固定在MCs上的肝细胞显示出比静态培养中的肝细胞显著更高的NH3代谢（p<0.01）。然而，单层培养的肝细胞比在静态培养中固定在MCs上的肝细胞显示出更高的葡萄糖合成（p<0.01）。总之，固定在MCs上的肝细胞被证明可作为混合人工肝中的生物反应器。
