A mini organ culture as a model for studying the gallbladder epithelium of mouse.
A mini organ culture of mouse gallbladder was developed as an alternative to primary cultures of epithelial cells of this organ. Small pieces of tissue were prepared and maintained in minimum essential Eagle medium with 10% foetal calf serum, for as long as 7 days. Qualitative and quantitative ultrastructural studies have been performed using electron microscopy. The viability of cells was evaluated by stereological quantification of endocytotic vesicles containing horseradish peroxidase and labelling of exocytotic glycoproteins with tannic acid. The morphology of tissue pieces during the 1st h of culturing and tissue isolated directly from animals exhibited no significant differences. However, after 4 h in culture degradative changes became evident in many cells. At that time, endo- and exocytosis were both dramatically reduced. After 24 h, the morphology, as well as endo- and exocytosis recovered and were comparable to the parameters of the tissue in vivo or after 1 h in culture. The endocytotic activity remained unchanged from day 1 to 7 of culturing, while the number of exocytotic vesicles gradually decreased after 2 days in culture. Our results prove that mini organ culture of gallbladder is morphologically and functionally comparable with the tissue in vivo and for studies of epithelium in culture it is more convenient than primary cultures.
Veranic
P
P
Institute of Cell Biology, Medical Faculty, Ljubljana, Slovenia.
Psenicnik
M
M
D000818:Animals; D004847:Epithelial Cells; D004848:Epithelium / Q000648:ultrastructure; D005260:Female; D005704:Gallbladder / Q000166:cytology*; D051379:Mice; D008854:Microscopy, Electron; D009924:Organ Culture Techniques / Q000379:methods*