J.Jpn. Surg. Soc.. 89(3): 388-393, 1988

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Original article

LONG-TERM MAINTENANCE OF HUMAN GALL-BLADDER EPITHELIA AS XENOGRAFTS FOLLOWING EXPLANT ORGAN CULTURE

The purpose of present study is to establish a method that should enable us to keep a human gall-bladder epithelial tissue alive for a period long enough to observe morphological change after an exposure to a certain carcinogenic agents. Human gall-bladder epithelia obtained from surgically resected specimen were kept as an explant organ culture in vitro for the first 2 weeks. They were, then, xenotransplanted to the subcutaneous (S.C.) space of the athymic nude mice. The gall-bladder tissues obtained from 7 cases of cholecystolithiasis and 1 case of cholecysto-choledocholithiasis were used as materials in the present study. And the longest survivor among them in the nude mice was extensively studied in the present paper. After an interval of 4 to 27 weeks, the xenografts were recovered from the recipient mice and studied by light microscopy and histochemistry. The morphological changes of the explant epithelia during the organ culture were also studied. Parts of the explants were implanted in 6 nude mice.
Each of the mice received 3 pieces of explants on the s.c. space in both of their flanks. All xenografts showed subcutaneous cyst formation. The cyst obtained 4 weeks after the implantation was consisted of a monolayer of cuboidal epithelial cells and those recovered from the recipient mice 24 and 27 weeks after the implantation were consisted of well growing collumnar and cuboidal epithelial cells.
In addition, sinus-like structures were observed beneath the cystic epithelia. The viability of the cells was excellent. Mucin production was evidenced by the positive staining by perioidic acid-Schiff-Alcian blue (PAS-AB) and high iron diamine-Alcian blue (HID-AB). The evidence shown here suggests a functional differentiation of the human gall-bladder epithelial cells that have regenerated in the explant organ culture in vitro. The potential applicability of the method used in the present paper to study human gall-bladder carcinogenesis as an experimental model is discussed.

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