Takahashi M, Matsue H, Matsushita M, Nakajima Y, Uchino J
First Department of Surgery, Hokkaido University School of Medicine, Sapporo, Japan.
Artif Organs. 1993 Jul;17(7):653-9. doi: 10.1111/j.1525-1594.1993.tb00610.x.
To use cultured human hepatocytes as a hybrid artificial liver, effective methods for isolating and culturing the hepatocytes from resected surgical specimens were investigated. Two different procedures for isolating hepatocytes, perfusion and agitation with a collagenase solution (Method 1) and perfusion with a mixed solution of collagenase and dispase (Method 2), were examined. The yield of isolated hepatocytes obtained by Method 2 (13.31 x 10(6) cells/g of liver) was significantly higher than that by Method 1 (0.94 x 10(6)). The warm ischemia time (0-90 min) of the liver fragments obtained did not disturb the viability and yield of the isolated hepatocytes. The gluconeogenesis and urea synthesis of the cultured human hepatocytes were well preserved for 10 days. These results show that for prolonged human hepatocyte culture (10 days), isolation from resected human liver tissues by a combination of the proteolytic enzymes collagenase and dispase was effective and warm ischemia was tolerated for up to 90 min, which indicates the possibility of using cultured human hepatocytes as a hybrid artificial liver.
为了将培养的人肝细胞用作混合人工肝，研究了从手术切除标本中分离和培养肝细胞的有效方法。考察了两种不同的肝细胞分离程序：用胶原酶溶液灌注和搅拌（方法1）以及用胶原酶和分散酶混合溶液灌注（方法2）。方法2获得的分离肝细胞产量（13.31×10⁶个细胞/克肝脏）显著高于方法1（0.94×10⁶个细胞/克肝脏）。所获得肝组织碎片的热缺血时间（0 - 90分钟）并未影响分离肝细胞的活力和产量。培养的人肝细胞的糖异生和尿素合成在10天内得到良好保存。这些结果表明，对于延长的人肝细胞培养（10天），通过蛋白水解酶胶原酶和分散酶联合从切除的人肝组织中分离是有效的，并且热缺血可耐受长达90分钟，这表明使用培养的人肝细胞作为混合人工肝的可能性。
