A comparison study of piglet hepatocyte culture in two types of artificial liver bioreactor

Objective To provide some new ideas and means for designing bioreactor by comparing two types of artificial liver bioreactor. Methods Microcarrier culture of piglet hepatocytes was performed in the culture bottle, after formation of microcarrier-spheroided aggregates the hepatocytes were seeded into extralumen of the bioreactor and cultured with a artificial capillary cell culture system. Then NH_4CL and Acetaminophen were added in the culture medium, the biological transforming function of the bioreactor was examined. The synthetic function of albumin of hepatocytes was tested, and the enzyme release of hepatocytes was examined. Viability of hepatocytes by both the trypan blue exclusion test and the ultrastructure of hepatocytes were determined. Results Hepatocytes which formed microcarrier-spheroided aggregates had some cell structures and reconstructed the cell-cell connections. Hepatocytes in the woven bioreactor maintained high viability within one week(viability of hepatocytes reached 75%±6.3% at 7 d). The bio-transformating function both for NH_4CL and Acetaminophen and synthetic function of albumin of hepatocytes were well maintained (0.57±0.13 mg/10~6 hepatocytes at 7 d), only a little of AST and LDH released from the cultured hepatocytes within one week. All these were much better than the common bioreactor(P0.05).Conclusion Hepatocyte-organoids having a tissue-like structure were equaly distributed into the extralumen of woven bioreactor under the support of woven hollow fiber which provide a three dimensions environment similar to that in vivo for growth and proliferation of hepatocytes. So it is a type of good performance bioreactor.