[Hybrid bioartificial liver using canine hepatocytes in primary culture].
Viable Hepatocytes were isolated from adult canine liver by in situ collagenase perfusion, and cultured on collagen coated borosilicate glass plates (100 X 200mm) at confluent cell density. The medium of hepatocytes in the primary culture was L-15 supplemented with aprotinin 5000U/L, proline 30mg/L, insulin 10(-8)M, dexamethasone 10(-8)M, glucagon 10(-8)M, and h-EGF 10ng/ml. Long-stroke type bioartificial liver module consisted of 200 glass plates with hepatocytes. It contained 6 billion primary cultured cells in total, that is almost equivalent to 30% of the normal canine liver. All hepatocytes in the module were quite viable during 2 weeks in the perfusion culture, and maintained various liver functions at a high level. Gluconeogenesis was 368.0 +/- 15.4mg/module/hr, albumin synthesis was 19.1 +/- 2.5mg/module/day, ureogenesis was 3.7 +/- 0.1mg/module/hr, and ammonia metabolism was 8.4mg/module/hr. Moreover, those functions were maintained at least 2 weeks in the canine plasma as well as in the culture medium with hormones. This hybrid bioartificial liver may exert various liver functions like a liver in situ.
Kumagai
F
F
First Department of Surgery, Hokkaido University School of Medicine, Sapporo, Japan.
D000418:Albumins / Q000096:biosynthesis; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D001187:Artificial Organs; D002478:Cells, Cultured; D003094:Collagen; D003470:Culture Media; D004285:Dogs; D005898:Glass; D005943:Gluconeogenesis; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology; D014508:Urea / Q000096:biosynthesis
D000418:Albumins; D003470:Culture Media; D000641:Ammonia; D014508:Urea; D003094:Collagen