Veranic P, Psenicnik M
Institute of Cell Biology, Medical Faculty, Ljubljana, Slovenia.
Biol Cell. 1996;88(3):145-51. doi: 10.1016/s0248-4900(97)83530-7.
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.
已开发出一种小鼠胆囊微型器官培养方法，作为该器官上皮细胞原代培养的替代方法。制备小块组织，并在含有10%胎牛血清的最低必需 Eagle 培养基中培养长达7天。使用电子显微镜进行了定性和定量超微结构研究。通过对含有辣根过氧化物酶的内吞小泡进行体视学定量以及用单宁酸标记外排糖蛋白来评估细胞活力。培养1小时期间的组织块形态与直接从动物分离的组织相比，无显著差异。然而，培养4小时后，许多细胞出现明显的降解变化。此时，内吞和外排作用均显著降低。24小时后，形态以及内吞和外排作用恢复，与体内组织或培养1小时后的组织参数相当。培养第1天至第7天，内吞活性保持不变，而培养2天后，外排小泡数量逐渐减少。我们的结果证明，胆囊微型器官培养在形态和功能上与体内组织相当，并且对于培养中的上皮研究而言，它比原代培养更方便。
