Uchino J, Matsue H, Takahashi M, Nakajima Y, Matsushita M, Hamada T, Hashimura E
First Department of Surgery, Hokkaido University School of Medicine, Sapporo, Japan.
ASAIO Trans. 1991 Jul-Sep;37(3):M337-8.
To produce a new hybrid artificial liver system, cultured pig hepatocytes were used in this study. Hepatocytes from normal healthy pigs were cultured in multiwell dishes. From the third day of cultivation, hepatocytes were divided into three groups according to the media used (Group 1, L-15; Group 2, normal human plasma; Group 3, plasma from hepatic failure [HF] patients). Measurements of cellular function of the cultured pig hepatocytes included metabolic activity (gluconeogenesis and ureogenesis), DNA content, and amino acid changes in the plasma. The ability to provide gluconeogenesis and ureogenesis by the cultured hepatocytes in HF plasma was maintained for 3 days, equal to that observed in Groups 1 and 2. DNA content was no different in the three groups. Elevated amino acid levels in the HF plasma, Phe, Met, Lys, and Gly, were significantly reduced by the cultured hepatocytes. The results indicate that the use of primary cultured pig hepatocytes is a step toward a hybrid artificial liver system, and a promising candidate as a bioreactor.
为制备一种新型杂交人工肝系统,本研究使用了培养的猪肝细胞。从正常健康猪获取的肝细胞在多孔培养皿中培养。从培养的第三天起,根据所用培养基将肝细胞分为三组(第1组,L-15;第2组,正常人血浆;第3组,肝衰竭[HF]患者的血浆)。对培养猪肝细胞的细胞功能测量包括代谢活性(糖异生作用和尿素生成)、DNA含量以及血浆中氨基酸变化。培养的肝细胞在HF血浆中提供糖异生作用和尿素生成的能力维持了3天,与第1组和第2组观察到的情况相当。三组中的DNA含量没有差异,但培养的肝细胞显著降低了HF血浆中升高的氨基酸水平,如苯丙氨酸、蛋氨酸、赖氨酸和甘氨酸水平降低。结果表明,使用原代培养猪肝细胞是迈向杂交人工肝系统的一步,并且作为生物反应器是一个有前景的候选者