[Purification and culture of mouse gallbladder epithelial cells in secondary culture using microexplant culture on collagen gel].

We established a new method of purification and culture of mouse gallbladder epithelial cells. In primary culture, mouse gallbladder tissue was cultured on collagen gel using microexplant culture method; the epithelial cells spread in a single sheet from the microexplant toward the periphery on the collagen gel. Tiny fragment of the collagen gel containing the border of epithelial cell layer was retransplanted onto the another collagen gel for secondary culture. This retransplantation allowed the proliferation of the epithelial cells without mesenchymal contamination, which spread on the collagen gel at the rate of 0.29 +/- 0.06 mm per day for more than two weeks. The cultured epithelial cells showed cuboid or columnar shape with focal mucus production, which is morphologically and functionally similar to the in vivo epithelial cells. This suggests that this in vitro culture method can be used to pathophysiological studies of biliary epithelial cells as well as for the method of purification of the epithelial cells.