Characterization of biliary epithelial cells isolated from needle biopsies of human liver in the presence of hepatocyte growth factor.
Current methods for the isolation of intrahepatic biliary epithelial cells from human liver rely upon relatively large segments of tissue, thereby limiting studies to cells isolated from patients with end-stage disease. To investigate a greater range of diseases and those at an earlier stage, we have developed a method to isolate biliary epithelial cells from biopsy-sized fragments of human liver. Tissue explants are cultured for > 4 weeks, and, in approximately 50% of samples incubated with medium containing hepatocyte growth factor, biliary epithelial cells begin to migrate from the fragments and proliferate. With time they form confluent pavements of cells that express cytokeratin 19 and gamma-glutamyl transpeptidase and are negative for markers of non-biliary cell phenotype. After subculturing, cells can be expanded, yielding substantial numbers for subsequent study in vitro. Cells can be isolated with a similar degree of success from adult normal liver, from a variety of liver diseases, and from post-transplant liver biopsies. Overall, pediatric tissue yielded cells less frequently than adult tissue. This novel technique is likely to have a major impact on the study of biliary pathophysiology, as small fragments of tissue removed from biopsies taken for diagnostic purposes can be used.
Strain
A J
AJ
Department of Biochemistry, University of Birmingham, United Kingdom.
Wallace
L
L
Joplin
R
R
Daikuhara
Y
Y
Ishii
T
T
Kelly
D A
DA
Neuberger
J M
JM
D000293:Adolescent; D000328:Adult; D000368:Aged; D001653:Bile Ducts, Intrahepatic / Q000473:pathology*; D001707:Biopsy, Needle; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003470:Culture Media / Q000737:chemistry; D003584:Cytological Techniques; D004848:Epithelium / Q000473:pathology; D005260:Female; D017228:Hepatocyte Growth Factor; D006801:Humans; D007223:Infant; D007381:Intermediate Filament Proteins / Q000032:analysis; D008107:Liver Diseases / Q000473:pathology* / Q000601:surgery; D016031:Liver Transplantation / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D013997:Time Factors
D003470:Culture Media; D007381:Intermediate Filament Proteins; D017228:Hepatocyte Growth Factor