Cultivation of primary porcine hepatocytes in an OXY-HFB for use as a bioartificial liver device.
Bioreactors being developed for bioartificial liver devices vary greatly in their construction. Until now, primary liver cells were cultivated either in sandwich configuration, as spheroids, or in special hollow fiber systems. Primary hepatocytes are demanding on their environment and have a high oxygen consumption. To get good results, optimal cultivation conditions are needed. The idea of the project was to investigate a new concept of an oxygenating hollow fiber bioreactor (OXY-HFB). The OXY-HFB should consist exclusively of oxygenating and internal heat exchange fibers to yield a simple and effective design. Primary liver cells were seeded on the surface of the fibers in the extrafiber space. Oxygen requirements and temperature control were supplied through the fibers. The culture medium was perfused through the extrafiber space and therefore brought into direct hepatocellular contact. The OXY-HFB concept offers different advantages. A high cell density of 2.5 x 10(7) cells/mL can be obtained. This results in a cell number of 2.5 x 10(9) liver cells per bioreactor. Furthermore, the OXY-HFB is easily handled because no incubator is required. To study the efficiency of this bioreactor technique, various parameters were investigated over a cultivation period of three weeks. These included urea synthesis, lactate formation, glucose elimination, albumin synthesis, oxygen level, and pH. Furthermore, the metabolites of diazepam were measured. The biochemical performance of the bioreactor remained stable over the investigated time period. These results demonstrate that porcine liver cells preserve their viability and primary metabolism in the OXY-HFB over the complete period of study.
Jasmund
Inka
I
Experimental Radiology, Hepatic Tissue Engineering, Medical School Hanover, Germany.
Langsch
Angelika
A
Simmoteit
Robert
R
Bader
Augustinus
A
D000818:Animals; D019149:Bioreactors; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D003975:Diazepam / Q000378:metabolism; D004867:Equipment Design; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism*; D019164:Liver, Artificial; D010100:Oxygen / Q000378:metabolism; D010477:Perfusion; D013552:Swine; D013997:Time Factors
D003470:Culture Media; D003975:Diazepam; D010100:Oxygen