Non-adhesive organ culture of human biliary epithelium with stroma.
OBJECTIVE
Explanted tissue has been shown to keep adult human cells in organ culture with a preserved morphology for at least one month as spheres in a non-adhesive organ culture. In the present study, we explored whether also human biliary epithelium can be grown in this manner, because the result may be of interest in studies of hepato-biliary-pancreatic carcinogenesis. MATERIAL AND METHODS. Small tissue samples were obtained from the gallbladder wall of patients who had been operated upon with cholecystectomy. Fragments of about 300 microm in diameter from each patient were cultured and investigated with light microscopy at the time of explantation and after 5, 10, 20, 30 and 40 days of culture. Scanning and transmission electron microscopy were performed to demonstrate the ultrastructure. Incubation of cultured fragments with the vital dyes revealed a viable epithelium.


RESULTS
At the time of explantation, all the tissue fragments had a rough appearance with an uneven, torn periphery, while during the first few days of culture they became rounder with a smooth-looking surface covering the entire circumference. This spheroid morphology persisted for the remainder of the culture period. The core of the fragments harboured connective tissue with vascular elements, fibroblasts and leucocytes. Immunostaining for cytokeratin 7, 19 and 20 revealed a strong positive staining of the epithelium.


CONCLUSIONS
These results show that biliary epithelium can be grown in vitro in a non-adhesive organ culture with their stroma.
Hoem
Dag
D
Department of Surgery, Institute of Surgical Sciences, Haukeland University Hospital, Bergen, Norway. Dag.Hoem@haukeland.no
Höstmark
Jens
J
Dalen
Helge
H
Andrén-Sandberg
Ake
A
D002470:Cell Survival; D053379:Chromogranin A / Q000032:analysis; D004848:Epithelium / Q000648:ultrastructure; D005704:Gallbladder / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D009924:Organ Culture Techniques; D018874:Spheroids, Cellular; D046509:Tissue Culture Techniques
D053379:Chromogranin A; D007633:Keratins