Chen Jyh-Ping, Yu Sheng-Chun, Hsu Brend Ray-Sea, Fu Shin-Huei, Liu Hwai-Shen
Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Kwei-San, Taoyuan 333, Taiwan, ROC.
Biotechnol Prog. 2003 Mar-Apr;19(2):522-7. doi: 10.1021/bp025720j.
Loofa sponge was investigated as a three-dimensional scaffold for stationary and perfusion culture of human hepatoblastoma cell line C3A/HepG2. In stationary culture, C3A/HepG2 cells in loofa cubes showed higher alpha-fetoprotein and albumin secretion rates than those in polyurethane foam (PU). To use loofa cylinders in a packed-bed reactor, immobilization of C3A/HepG2 cells by recirculating medium at 26 mL/min (superficial velocity = 51.7 cm/min) resulted in a cell loading density of 5.15 x 10(7) cells/cm(3)-loofa. This cell loading density is higher than values reported in the literature for packed-bed reactor intended for bioartificial liver. During 9 days of perfusion culture in the reactor, immobilized C3A/HepG2 showed steady synthesis of albumin with an average synthesis rate at 42.2 microg/10(6) cells/day. These experimental results and observations by SEM suggested that loofa sponge is a suitable scaffold for high-density culture of human hepatocyte cell line and the immobilized cells could express high levels of liver-specific functions.
研究了丝瓜海绵作为人肝癌细胞系C3A/HepG2固定化培养和灌注培养的三维支架。在固定化培养中，丝瓜方块中的C3A/HepG2细胞比聚氨酯泡沫（PU）中的细胞表现出更高的甲胎蛋白和白蛋白分泌率。为了在填充床反应器中使用丝瓜圆柱，以26 mL/min的流速（表观流速 = 51.7 cm/min）循环培养基固定C3A/HepG2细胞，得到的细胞负载密度为5.15×10⁷个细胞/cm³丝瓜。该细胞负载密度高于文献报道的用于生物人工肝的填充床反应器的数值。在反应器中进行9天的灌注培养期间，固定化的C3A/HepG2细胞白蛋白合成稳定，平均合成速率为42.2 μg/10⁶个细胞/天。这些实验结果和扫描电镜观察表明，丝瓜海绵是适合人肝细胞系高密度培养的支架，且固定化细胞能够高水平表达肝脏特异性功能。
