Wang Ying-jie, Liu Hong-ling, Guo Hai-tao, Liu Jun, Wen Hong-wei, Wang Yu-ming
Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Zhonghua Gan Zang Bing Za Zhi. 2003 Jun;11(6):358-60.
To study the method of preparing the hollow fiber bioreactor for culturing pig hepatocytes.
Hepatocytes were isolated from experimental suckling minipigs by two-step perfusion with collagenase, and seeded onto hollow fiber bioreactor, then cultured with an artificial capillary cell culture system. The albumin-excretion, lidocaine-transforming rate, lactate dehydrogenase (LDH) release and the cell viability in bioreactors were examined.
The porcine albumin could be detected by SDS/PAGE on the 2nd, 4th, 6th day. The rates of lidocaine-transforming ranged from 89.6% to 96.1%. The release of LDH into the culture medium increased from (23.7+/-4.6) U/L to (127.8+/-17.4) U/L (F=39.582, P<0.01) during the experiments, and the viability of pig hepatocytes in hollow fiber bioreactor reduced from 95.8%+/-0.3% to 83.8%+/-4.7% (t=5.135, P<0.01).
The hollow fiber bioreactor for culturing pig hepatocytes can be prepared by artificial capillary cell culture system, which provides a certain liver-specific function in 1 week.
研究制备用于培养猪肝细胞的中空纤维生物反应器的方法。
采用两步胶原酶灌注法从实验用哺乳仔猪分离肝细胞，接种于中空纤维生物反应器，然后用人工毛细管细胞培养系统进行培养。检测生物反应器中白蛋白分泌、利多卡因转化率、乳酸脱氢酶（LDH）释放及细胞活力。
在第2、4、6天通过SDS/PAGE可检测到猪白蛋白。利多卡因转化率在89.6%至96.1%之间。实验期间，培养基中LDH释放量从（23.7±4.6）U/L增加到（127.8±17.4）U/L（F=39.582，P<0.01），中空纤维生物反应器中猪肝细胞活力从95.8%±0.3%降至83.8%±4.7%（t=5.135，P<0.01）。
采用人工毛细管细胞培养系统可制备用于培养猪肝细胞的中空纤维生物反应器，其在1周内具有一定的肝脏特异性功能。
