[Preparation of hollow fiber bioreactor for culturing pig hepatocytes].
Yingjie Wang, Hongling Liu, Haitao Guo, Jun Liu, Hong-wei Wen, Yuming Wang
Abstract
Yingjie Wang, Hongling Liu, Haitao Guo, Jun Liu, Hong-wei Wen, Yuming Wang
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.
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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.
Key concepts: Bioreactor, Bioartificial liver device, Collagenase, Lactate dehydrogenase, Albumin, Hepatocyte, Chromatography, Fiber