title: A Modified Method for Purifying Gallbladder Epithelial Cells Using Fluorescence-activated Cell Sorting.
abstract: 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.
  
  
  
  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).
  
  
  
  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 0.82×106pure BECs that survived were obtained and seeded on a collagen gel plate. However, these pure BECs showed almost no proliferation.
  
  
  
  Pure BECs could be accumulated using FACS. However, the number of BECs was insufficient for the culturing process.
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Imamura
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Adachi
authors.affiliation: Department of Surgery, National Hospital Organization Nagasaki Medical Center, Omura, Japan.
authors.forename: Amane
authors.initials: A
authors.lastname: Kitasato
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sakai
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Ono
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Takanobu
authors.initials: T
authors.lastname: Hara
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Natsuda
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Soyama
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Hidaka
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
authors.forename: Mitsuhisa
authors.initials: M
authors.lastname: Takatsuki
authors.affiliation: Department of Surgery, National Hospital Organization Nagasaki Medical Center, Omura, Japan.
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Kuroki
authors.affiliation: Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan sueguchi@nagasaki-u.ac.jp.
authors.forename: Susumu
authors.initials: S
authors.lastname: Eguchi
mesh_terms: D000818:Animals; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D003609:Dactinomycin / Q000031:analogs & derivatives / Q000378:metabolism; D000071858:Epithelial Cell Adhesion Molecule / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005704:Gallbladder / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008810:Mice, Inbred C57BL; D008858:Microscopy, Phase-Contrast; D015203:Reproducibility of Results
chemical_list: D000071858:Epithelial Cell Adhesion Molecule; D003609:Dactinomycin; C025942:7-aminoactinomycin D
keywords: Biliary epithelial cell; fluorescence-activated cell sorting; isolation; mouse gallbladder