Characterization of human hepatocyte lines derived from normal liver tissue.
Four separate continuous lines of human hepatocytes (HH01, HH02, HH09, HH25) were developed from normal liver tissue by subjecting cocultures of human hepatocytes with rat liver epithelial cells in a highly enriched medium to frequent subculturing. The addition of conditioned medium from either the human hepatoma line Hep G2 or one of these stable human hepatocyte lines (HH09) appeared to facilitate establishment of line HH25. These human hepatocyte lines have been in continuous culture for 2 to 5 yr and consist of approximately 95% human cells by analysis of cell surface antigens. Cytogenetic analysis also confirmed the human origin of these cells and showed clonal origin with abnormal ploidy. Cells in these human hepatocyte lines retain morphological features of hepatocytes by both light and electron microscopy. They also retain glucose-6-phosphatase activity and secrete proteins characteristic of hepatocytes, such as albumin, alpha-fetoprotein and transferrin. After incubation with 13 mumol/L dibenz(a,h) anthracene for 24 hr, each line had detectable activity of aryl hydrocarbon hydroxylase, ethoxyresorufin O-deethylase and methoxyresorufin O-demethylase. Thus, these human hepatocyte lines retain important differentiated characteristics of hepatocytes. Derived from normal liver tissue, they appear to be immortalized. They provide a new model system for studying human hepatocellular drug metabolism. These lines may also be useful for studying the regulation of synthesis of albumin, alpha-fetoprotein and other proteins in human hepatocytes, determining the effects of cytokines and growth factors and designing systems to effect gene transfer into human hepatocytes for the purpose of gene therapy.
Roberts
E A
EA
Division of Gastroenterology, Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
Letarte
M
M
Squire
J
J
Yang
S
S
D000418:Albumins / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D002648:Child; D002877:Chromosomes, Human; D003470:Culture Media; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005260:Female; D005952:Glucose-6-Phosphatase / Q000032:analysis; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis; D006801:Humans; D007223:Infant; D016023:Integrins / Q000096:biosynthesis; D008099:Liver / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D010088:Oxidoreductases / Q000378:metabolism; D051381:Rats; D014168:Transferrin / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
D000418:Albumins; D003470:Culture Media; D015395:Histocompatibility Antigens Class I; D016023:Integrins; D014168:Transferrin; D000509:alpha-Fetoproteins; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C076307:methoxyresorufin-O-demethylase; D001189:Aryl Hydrocarbon Hydroxylases; D019363:Cytochrome P-450 CYP1A1; D005952:Glucose-6-Phosphatase