J.Jpn. Surg. Soc.. 90(4): 546-555, 1989

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Original article

HYBRID BIOARTIFICAL 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 (100×200mm) at confluent cell density. The medium of hepatocytes in the primary culture was L-15 supplemented with aprotinin 5000U/L, prolin 30mg/L, insulin 10^-8M, dexamethasone 10^-8M, glucagon 10^-8M, 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.

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