Long-term culture of hepatocytes from human adults
Journal
Journal of Biomedical Science
Journal Volume
5
Journal Issue
6
Pages
435-440
Date Issued
1998
Author(s)
Abstract
A long-term primary human hepatocyte culture retraining liver-specific functions is important and essential for basic research and for the future development of hepatocyte-based applications. We established a normal hepatocyte culture system from excess normal tissues obtained from adult liver cancer patients who received partial liver resection. Hepatocytes were isolated after perfusion and enzymatic disaggregation, and were first maintained in hormonally, defined media on a Matrigel matrix, and then transferred to collagen sandwich gel. The hepatocytes formed clusters on the Matrigel matrix and increased in size and numbers with time of culture and eventually grew into spheroids of variable sizes. After being transferred to collagen gel, the cells migrated outward from spheroids to form a monolayer with cuboidal or polygonal cell shapes with granular cytoplasm and continued to proliferate. Cellular functions specific for hepatocytes were analyzed using immunoblot assay for proteins specifically secreted by the liver cells on different days of culture. The cells secreted albumin, transferrin and α- fetoprotein consistently for more than 100 days, to a maximum of 150 days. Thus, we have established a long-term culture of hepatocytes from human adults, which will be useful for basic studies of liver physiology such as metabolism and morphogenesis, as well as for other applications in the study of infectious hepatitis, pharmacology, pharmacokinetics, and toxicology.
Subjects
Albumin; Collagen gel sandwich; Human hepatocyte; Matrigel matrix; Primary culture; Transferrin; α-Fetoprotein
SDGs
Other Subjects
albumin; alpha fetoprotein; matrigel; transferrin; article; cell culture; cell isolation; controlled study; human; human cell; liver carcinoma; liver cell; liver metabolism; morphogenesis; priority journal
Publisher
BioMed Central Ltd.
Type
journal article