Original Articles
Copyright ©The Author(s) 2000. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 15, 2000; 6(3): 365-370
Published online Jun 15, 2000. doi: 10.3748/wjg.v6.i3.365
Primary porcine hepatocytes with portal vein serum cultured on microcarriers or in spheroidal aggregates
Yi Gao, Huan-Zhang Hu, Ji-Zhen Yang, Ke Chen
Yi Gao, Huan-Zhang Hu, Ke Chen and Ji-Zhen Yang, Department of Hepatobiliary Surgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, Guangdong Province, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Science Foundation of China, No. 39570212
Correspondence to: Dr. Yi Gao, Department of Hepatobiliary Surgery, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, Guangdong Province, China
Telephone: +86-20-85143556
Received: January 10, 2000
Revised: February 3, 2000
Accepted: March 2, 2000
Published online: June 15, 2000
Abstract

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.

Keywords: porcine hepatocytes; microcarriers; cell culture; spheroidal aggregate culture; portal vein serum