用于培养猪肝细胞的中空纤维生物反应器的制备

[Preparation of hollow fiber bioreactor for culturing pig hepatocytes].

作者信息

Wang Ying-jie, Liu Hong-ling, Guo Hai-tao, Liu Jun, Wen Hong-wei, Wang Yu-ming

机构信息

Institute of Infectious Diseases, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2003 Jun;11(6):358-60.

PMID:12837217

Abstract

OBJECTIVE

To study the method of preparing the hollow fiber bioreactor for culturing pig hepatocytes.

METHODS

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.

RESULTS

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).

CONCLUSION

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.

摘要

目的

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。

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。

结果

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）。