Efficacy of a polyurethane foam/spheroid artificial liver by using human hepatoblastoma cell line (Hep G2)
Efficacy of a polyurethane foam/spheroid artificial liver by using human hepatoblastoma cell line (Hep G2)
复制标题
DOI：
10.3727/000000003783985151
复制
发表时间：
2003-01-01
影响因子：
3.3
通讯作者：
Funatsu, K
中科院分区：
文献类型：
--
作者：
Fukuda, J;Okamura, K;Funatsu, K
我们研究了人肝母细胞瘤细胞系（Hep G2）与人原代肝细胞（HH）和猪原代肝细胞（PH）作为混合型人工肝支持系统（HALSS）细胞来源的可用性。所有三种肝细胞自发形成球形的多细胞聚集体（球体）的100-200微米直径的孔的PUF内3天的文化。在PUF静止培养中，Hep G2球状体恢复了单层培养中失去的氨去除活性，尽管每个单位细胞数的去除约为HH球状体的十分之一和约PH球状体的八分之一。PUF球状体培养也能显著提高Hep G2各单位细胞白蛋白和纤维蛋白原的合成活性，其合成活性约为单层培养的2倍。Hep G2球状体的白蛋白分泌活性与PH球状体基本相同。在本实验中，HH几乎不分泌这些蛋白质，可能是因为它们是在无血清培养基中培养的。在循环培养的PUF模块中，HH具有类似于静止培养的高氨去除和低合成活性。Hep G2增殖到高细胞密度，例如在培养10天时约为4.8 × 107个细胞/cm 3-模块。虽然Hep G2球状体在每个细胞中具有低的氨去除活性，但是由于通过细胞增殖进行的高细胞密度培养，因此在培养7天时PUF模块中的去除率与PH几乎相同。在PUF模块中Hep G2的白蛋白分泌速率也随着细胞增殖而增加，并且在培养7天时比PH的初始速率高约10倍。这些结果表明Hep G2是PUF-HALSS的潜在细胞来源。
We investigated the availability of human hepatoblastoma cell line (Hep G2), compared with human primary hepatocytes (HH) and porcine primary hepatocytes (PH), as a cell source for the hybrid artificial liver support system (HALSS) by using polyurethane foam (PUF). All three kinds of hepatocytes spontaneously formed spherical multicellular aggregates (spheroids) of 100-200 mum diameter in the pores of PUF within 3 days of culture. In a PUF stationary culture, Hep G2 spheroids recovered the ammonia removal activity that was lost in monolayer culture, although the removal for each unit cell number was about one tenth that of HH spheroids and about one eighth of PH spheroids. The synthesis activities of albumin and fibrinogen of each unit cell number of Hep G2 were also upregulated by PUF spheroid culture, and were about twice as high as in monolayer culture. The albumin secretion activity of Hep G2 spheroids was almost the same as that of PH spheroids. HH scarcely secreted these proteins in this experiment, probably because they were cultured in a serum-free medium. In the PUF module in a circulation culture, HH had high ammonia removal and low synthesis activities similar to stationary culture. Hep G2 proliferated to a high cell density, such as about 4.8 x 10(7) cells/cm(3) -module at 10 days of culture. Although Hep G2 spheroids had low ammonia removal activity in each cell, the removal rate in the PUF module was almost the same as for PH at 7 days of culture because of the high cell density culture by cell proliferation. The albumin secretion rate by Hep G2 in the PUF module also increased with cell proliferation and was about 10 times higher than the initial rate for PH at 7 days of culture. These results suggest that Hep G2 is a potential cell source for the PUF-HALSS.