A Modified Method for Purifying Gallbladder Epithelial Cells Using Fluorescence-activated Cell Sorting
A Modified Method for Purifying Gallbladder Epithelial Cells Using Fluorescence-activated Cell Sorting
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DOI：
10.21873/invivo.11041
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发表时间：
2017-03-01
期刊：
影响因子：
2.3
通讯作者：
Eguchi, Susumu
中科院分区：
文献类型：
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作者：
Imamura, Hajime;Adachi, Tomohiko;Eguchi, Susumu
背景：我们以前报道了一种分离和培养胆管上皮细胞（BEC）的方法。本研究的目的是重新考虑使用小鼠胆囊获得纯BEC的方法。材料与方法：通过荧光激活细胞分选（FACS）分选仅从胆囊获得的细胞，用于基于上皮细胞粘附分子（EpCAM）的表达进行纯化。根据7-氨基放线菌素D（7-AAD）的阴性表达来测量存活率。结果：经鉴定，75%以上的胆囊BEC为纯BEC。EpCAM分析显示73.3%的细胞为7-AAD阴性。最后，获得存活的0.82 × 10（6）纯BEC并接种在胶原凝胶板上。然而，这些纯BEC几乎没有显示增殖。结论：流式细胞仪可获得纯度较高的BEC。然而，BEC的数量不足以用于培养过程。
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 0.82x10(6) 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.