Demonstration and maintenance of mucus secretion in cultured human gallbladder epithelial cells |
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Authors: | Soichi Yoshitomi Kohji Miyazaki Fumio Nakayama |
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Institution: | (1) Department of Surgery I, Kyushu University Faculty of Medicine, 812 Fukuoka, Japan |
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Abstract: | Summary The method of human gallbladder epithelial cell culture has been developed successfully with active mucus secretory function.
Human gallbladder epithelial cells were dissociated by Dispase digestion from the specimens obtained by cholecystectomy for
uncomplicated gallbladder stone cases. The dissociated cells formed a monolayer in Eagle’fs minimum essential medium supplemented
with 10% fetal bovine serum within 24 h after the inoculation. These cells were maintained for at least 2 wk without fibroblastic
overgrowth. Cultured cells contained periodic acid Schiff-positive material in cellular cytoplasm for 3 d. On transmission
electron microscopy these materials were identified as mucous secretory granules. Mucous secretory function was determined
by 3H]glucosamine incorporation. Sixty percent of the secreted glycoproteins labeled with 3H]glucosamine was eluted in excluded fractions of Sepharose 4B gel filtration, which were considered to be mucous glycoprotein,
because they were found to be resistant to proteoglycan-specific enzymes such as hyaluronidase, chondroitinase ABC, heparitinase,
and heparinase. The mucous glycoprotein secretion was maintained for 3 d and found to be inhibited in a dose-dependent manner
by monensin (10−7 to 10−5
M) which is a known blocker of secretory function. |
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Keywords: | human gallbladder epithelium primary culture mucus secretion ultrastructure monensin |
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