Department of Biomedical Engineering, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan.
J Biotechnol. 2010 Aug 2;148(4):226-32. doi: 10.1016/j.jbiotec.2010.06.002. Epub 2010 Jun 9.
To develop a tissue-engineered bioartificial liver (BAL), perfusion cultures of mouse and pig fetal liver cells (FLCs) immobilized within a three-dimensional (3D) porous scaffold were performed utilizing a packed-bed reactor system. These FLCs were cultured under different medium flow rate conditions, and the effects of the flow rates on cell growth and the hepatic functions of the FLCs were investigated. In the cultures of mouse FLCs, the medium flow suppressed cell growth and the albumin secretion activity of the FLCs, and considerably lower albumin secretion than that in the 3D stationary control cultures was obtained in the perfusion cultures. In the case of pig FLCs, cell growth was also inhibited by the medium flow, however, the cells exhibited higher tolerance to the medium flow compared with mouse FLCs. The albumin secretion activity of pig FLCs was well maintained under an extremely low flow rate condition (4.8 mm/min in the reactor), and activity higher than the 3D stationary cultures was detected at a later stage (after 20 days in the perfusion cultures). These results revealed that FLCs are quite sensitive to medium flow and an extremely low shear stress is required for the perfusion cultures of FLCs.
为了开发组织工程化的生物人工肝（BAL），我们使用填充床反应器系统进行了固定在三维（3D）多孔支架内的小鼠和猪胎肝细胞（FLC）的灌注培养。这些 FLC 是在不同的培养基流速条件下培养的，研究了流速对细胞生长和 FLC 肝功能的影响。在小鼠 FLC 的培养中，培养基流速抑制了细胞生长和 FLC 的白蛋白分泌活性，并且在灌注培养中获得的白蛋白分泌明显低于 3D 静态对照培养。对于猪 FLC，细胞生长也受到培养基流动的抑制，但与小鼠 FLC 相比，细胞对培养基流动的耐受性更高。在极低的流速条件下（反应器中为 4.8mm/min），猪 FLC 的白蛋白分泌活性得到很好的维持，并且在灌注培养的后期（20 天后）检测到比 3D 静态培养更高的活性。这些结果表明 FLC 对培养基流动非常敏感，并且 FLC 的灌注培养需要极低的剪切应力。
