Published online Jun 15, 2000. doi: 10.3748/wjg.v6.i3.365
Revised: February 3, 2000
Accepted: March 2, 2000
Published online: June 15, 2000
AIM: To develop a culture mode providing durable biomaterials with high yields and activities used in bioartificial liver.
METHODS: Hepatocytes were isolated from a whole pig liver by Seglen′s method of orthotopic perfusion with collagenase. In culture on microcarriers, primary porcine hepatocytes were inoculated at a concentration of 5 × 107/mL into the static culture systems containing 2 g/L Cytodex-3, then supplemented with 100 mL/L fetal calf serum (FCS) or 100 mL/L porcine portal vein serum (PPVS) respectively. In spheroidal aggregate culture hepatocytes were inoculated into 100 mL siliconized flasks at a concentrati on of 5.0 × 106/mL.
RESULTS: In culture on microcarriers hepatocytes tended to aggregate on Cytodex-3 obviously after being inoculated. Typical multi-cellular aggregated spheroids could be found in the two systems 24-48 h after hepatocytes were cultured. The morphological charact-eristics and synthetic functions were maintained for 5 wk in FCS culture system and 8 wk in PPVS culture system. In spheroidal aggregate culture about 80%-90% isolated hepatocytes became aggregated spheroids 24 h after cultured in suspension and mean diameter of the spheroids was 100 μm. The relationship among the hepatocytes resembled that in the liver in vivo. Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytes cultured on monolayers.
CONCLUSION: As high-yields and high-activity modes of culture on microcarriers or in spheroidal aggregate culture with portal vein serum are promising to provide biomaterials for bioartificial liver (BAL) efficiently.