1 Department of Nutritional Science and Food Safety, Faculty of Applied Bioscience, Tokyo University of Agriculture , Tokyo, Japan .
2 Department of Developmental Biology, Washington University School of Medicine in St. Louis , St. Louis, Missouri.
Tissue Eng Part C Methods. 2018 Aug;24(8):480-485. doi: 10.1089/ten.TEC.2018.0127.
Proper identification of pancreatic ducts is a major challenge for researchers performing partial duct ligation (PDL), because pancreatic ducts, which are covered with acinar cells, are translucent and thin. Although damage to pancreatic ducts may activate quiescent ductal stem cells, which may allow further investigation into ductal stem cells for therapeutic use, there is a lack of effective techniques to visualize pancreatic ducts. In this study, we report a new method for identifying pancreatic ducts. First, we aimed to visualize pancreatic ducts using black, waterproof fountain pen ink. We injected the ink into pancreatic ducts through the bile duct. The flow of ink was observed in pancreatic ducts, revealing their precise architecture. Next, to visualize pancreatic ducts in live animals, we injected fluorescein-labeled bile acid, cholyl-lysyl-fluorescein into the mouse tail vein. The fluorescent probe clearly marked not only the bile duct but also pancreatic ducts when observed with a fluorescent microscope. To confirm whether the pancreatic duct labeling was successful, we performed PDL on Neurogenin3 (Ngn3)-GFP transgenic mice. As a result, acinar tissue is lost. PDL tail pancreas becomes translucent almost completely devoid of acinar cells. Furthermore, strong activation of Ngn3 expression was observed in the ligated part of the adult mouse pancreas at 7 days after PDL.
正确识别胰腺导管是进行部分导管结扎(PDL)的研究人员面临的主要挑战,因为胰腺导管被腺泡细胞覆盖,呈半透明且很薄。尽管导管损伤可能会激活静止的导管干细胞,这可能有助于进一步研究导管干细胞用于治疗用途,但目前缺乏有效的技术来可视化胰腺导管。在这项研究中,我们报告了一种识别胰腺导管的新方法。首先,我们旨在使用黑色防水钢笔墨水可视化胰腺导管。我们通过胆管将墨水注入胰腺导管。观察到墨水在胰腺导管中流动,揭示了其精确的结构。接下来,为了在活体动物中可视化胰腺导管,我们将荧光素标记的胆汁酸、胆酰-赖氨酸-荧光素注入小鼠尾静脉。当用荧光显微镜观察时,荧光探针不仅清晰地标记了胆管,还标记了胰腺导管。为了确认胰腺导管标记是否成功,我们对 Neurogenin3 (Ngn3)-GFP 转基因小鼠进行了 PDL。结果,腺泡组织丢失。PDL 后的第 7 天,结扎部分的胰腺尾变得几乎完全透明,几乎没有腺泡细胞。此外,在 PDL 后 7 天,成年小鼠胰腺的结扎部位观察到强烈的 Ngn3 表达激活。