International Center for Biotechnology, Osaka University, 2-1, Yamada-oka, Suita, Osaka, 565, Japan.
Cytotechnology. 1997 May;24(1):39-45. doi: 10.1023/A:1007927906986.
Selection of a cell line suitable for a hybrid artificial liver model employing cellulose porous beads (CPBs) was investigated. Hep G2 cells grown in a culture dish exhibited appreciably higher ureogenesis and gluconeogenesis activities than those grown in CPBs. SEM observation of CPBs revealed marked difference in the distribution of attached cells from one bead to another, and showed that almost all the cell-bearing micropores were completely packed with cells.With the aim of selecting a cell line not prone to excessive aggregation and which grows moderately so as not to fill up the micropores, cells of 6 cell lines, HLE, HLF, Hep 3B, PLC/PRF/5, Huh 7 and Hep G2, were cultivated in dishes. Hep G2, HLE, and HLF increased to 5 × 10(5) cells/cm(2), whereas PLC/PRF/5 grew only to 5 × 10(4), and Hep 3B and Huh 7 up to 2 × 10(4) cells/cm(2). The specific activities of ureogenesis and gluconeogenesis of Huh 7 were the highest among the lines tested - 42- and 7-fold those of Hep G2, respectively. When the 6 cell lines were grown in a submerged culture with 0.6 g/l of CPBs, Huh 7 had the lowest cell concentration of 0.54 × 10(6) cells/ml, and the highest activities of ammonia consumption and urea and glucose production (1.38 μ mol NH(3), 99 nmol urea, and 14.5 nmol glucose/10(6)cells/h). Consequently, Huh 7 is considered to be a suitable cell line for use in the development of an artificial liver model employing porous beads.
研究了适合使用纤维素多孔珠（CPBs）的杂交人工肝模型的细胞系选择。与在 CPBs 中生长的细胞相比，在培养皿中生长的 Hep G2 细胞表现出明显更高的尿素生成和糖异生活性。对 CPBs 的 SEM 观察显示，附着细胞在一个珠粒到另一个珠粒之间的分布有明显差异，并且表明几乎所有带细胞的微孔都完全被细胞填充。为了选择不易过度聚集且生长适中的细胞系，以使微孔不被填满，将 6 种细胞系（HLE、HLF、Hep 3B、PLC/PRF/5、Huh 7 和 Hep G2）的细胞在培养皿中进行培养。Hep G2、HLE 和 HLF 增加到 5×10(5)个细胞/cm(2)，而 PLC/PRF/5 仅增加到 5×10(4)个细胞/cm(2)，Hep 3B 和 Huh 7 增加到 2×10(4)个细胞/cm(2)。在测试的细胞系中，Huh 7 的尿素生成和糖异生的比活性最高 - 分别为 Hep G2 的 42 倍和 7 倍。当 6 种细胞系在 0.6 g/l 的 CPBs 中进行浸没培养时，Huh 7 的细胞浓度最低，为 0.54×10(6)个细胞/ml，氨消耗和尿素和葡萄糖生成的活性最高（1.38 μmol NH(3)、99 nmol 尿素和 14.5 nmol 葡萄糖/10(6)细胞/h）。因此，Huh 7 被认为是一种适合用于开发使用多孔珠的人工肝模型的细胞系。
