一种使用荧光激活细胞分选技术纯化胆囊上皮细胞的改良方法。
A Modified Method for Purifying Gallbladder Epithelial Cells Using Fluorescence-activated Cell Sorting.
作者信息
Imamura Hajime, Adachi Tomohiko, Kitasato Amane, Sakai Yusuke, Ono Shinichiro, Hara Takanobu, Natsuda Koji, Soyama Akihiko, Hidaka Masaaki, Takatsuki Mitsuhisa, Kuroki Tamotsu, Eguchi Susumu
机构信息
Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Department of Surgery, National Hospital Organization Nagasaki Medical Center, Omura, Japan.
出版信息
In Vivo. 2017 Mar-Apr;31(2):169-173. doi: 10.21873/invivo.11041.
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 0.82×10 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.
背景
我们之前报道了一种分离和培养胆管上皮细胞（BECs）的方法。本研究的目的是重新考虑使用小鼠胆囊获取纯BECs的方法。
材料与方法
仅从胆囊获得的细胞通过荧光激活细胞分选（FACS）进行分选，以基于上皮细胞粘附分子（EpCAM）的表达进行纯化。基于7-氨基放线菌素D（7-AAD）的阴性表达来测量存活率。
结果
超过75%的胆囊细胞被确定为纯BECs。对EpCAM的分析显示，73.3%的细胞为7-AAD阴性。最后，获得了存活的0.82×10个纯BECs并接种在胶原凝胶板上。然而，这些纯BECs几乎没有增殖。
结论
可以使用FACS积累纯BECs。然而，BECs的数量不足以用于培养过程。
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