BACKGROUND: We have previously reported a procedure for isolating and culturing biliary epithelial cells (BECs). The aim of this study was to reconsider the method for obtaining pure BECs using the mouse gallbladder. MATERIALS AND METHODS: Cells that were obtained from the gallbladder alone were sorted by fluorescence-activated cell sorting (FACS) for purifying based on the expression of the epithelial cell adhesion molecule (EpCAM). The viability rate was measured based on the negative expression of 7-aminoactinomycin D (7-AAD). RESULTS: More than 75% of cells from the gallbladder were determined to be pure BECs. An analysis of the EpCAM revealed that 73.3% of the cells were 7-AAD-negative. Finally, the 0.82×106 pure BECs that survived were obtained and seeded on a collagen gel plate. However, these pure BECs showed almost no proliferation. CONCLUSION: Pure BECs could be accumulated using FACS. However, the number of BECs was insufficient for the culturing process. Copyright
BACKGROUND: We have previously reported a procedure for isolating and culturing biliary epithelial cells (BECs). The aim of this study was to reconsider the method for obtaining pure BECs using the mouse gallbladder. MATERIALS AND METHODS: Cells that were obtained from the gallbladder alone were sorted by fluorescence-activated cell sorting (FACS) for purifying based on the expression of the epithelial cell adhesion molecule (EpCAM). The viability rate was measured based on the negative expression of 7-aminoactinomycin D (7-AAD). RESULTS: More than 75% of cells from the gallbladder were determined to be pure BECs. An analysis of the EpCAM revealed that 73.3% of the cells were 7-AAD-negative. Finally, the 0.82×106 pure BECs that survived were obtained and seeded on a collagen gel plate. However, these pure BECs showed almost no proliferation. CONCLUSION: Pure BECs could be accumulated using FACS. However, the number of BECs was insufficient for the culturing process. Copyright
Authors: Piero Dalerba; Tomer Kalisky; Debashis Sahoo; Pradeep S Rajendran; Michael E Rothenberg; Anne A Leyrat; Sopheak Sim; Jennifer Okamoto; Darius M Johnston; Dalong Qian; Maider Zabala; Janet Bueno; Norma F Neff; Jianbin Wang; Andrew A Shelton; Brendan Visser; Shigeo Hisamori; Yohei Shimono; Marc van de Wetering; Hans Clevers; Michael F Clarke; Stephen R Quake Journal: Nat Biotechnol Date: 2011-11-13 Impact factor: 54.908