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Mouse organoid culture is a suitable model to study esophageal ion transport mechanisms
American Journal of Physiology-Cell Physiology ( IF 5.5 ) Pub Date : 2021-09-15 , DOI: 10.1152/ajpcell.00295.2021
Marietta Margaréta Korsós 1 , Tamás Bellák 2, 3 , Eszter Becskeházi 1 , Eleonóra Gál 1 , Zoltán Veréb 4 , Péter Hegyi 5, 6, 7 , Viktória Venglovecz 1
Affiliation  

Altered esophageal ion transport mechanisms play a key role in inflammatory and cancerous diseases of the esophagus, but epithelial ion processes have been less studied in the esophagus because of the lack of a suitable experimental model. In this study, we generated 3D esophageal organoids (EOs) from two different mouse strains and characterized the ion transport processes of the EOs. EOs form a cell-filled structure with a diameter of 250-300 µm and generated from epithelial stem cells as shown by FACS analysis. Using conventional PCR and immunostaining, the presence of Slc26a6 Cl/HCO3 anion exchanger (AE), Na+/H+ exchanger (NHE), Na+/HCO3- cotransporter (NBC), cystic fibrosis transmembrane conductance regulator (CFTR) and anoctamin 1 Cl channels were detected in EOs. Microfluorimetric techniques revealed high NHE, AE, and NBC activities, whereas that of CFTR was relatively low. In addition, inhibition of CFTR led to functional interactions between the major acid-base transporters and CFTR. We conclude that EOs provide a relevant and suitable model system for studying the ion transport mechanisms of esophageal epithelial cells, and they can be also used as preclinical tools to assess the effectiveness of novel therapeutic compounds in esophageal diseases associated with altered ion transport processes.

中文翻译:

小鼠类器官培养是研究食管离子转运机制的合适模型

改变的食管离子转运机制在食管的炎症和癌性疾病中起关键作用,但由于缺乏合适的实验模型,食管上皮离子过程的研究较少。在这项研究中,我们从两种不同的小鼠品系中生成了 3D 食管类器官 (EOs),并表征了 EOs 的离子转运过程。如 FACS 分析所示,EO 形成直径为 250-300 µm 的细胞填充结构,由上皮干细胞生成。使用常规 PCR 和免疫染色,Slc26a6 Cl - /HCO 3 -阴离子交换剂 (AE)、Na + /H +交换剂 (NHE)、Na + /HCO 3 -的存在在EOs中检测到协同转运蛋白(NBC)、囊性纤维化跨膜电导调节剂(CFTR)和anoctamin 1 Cl -通道。微荧光技术显示高 NHE、AE 和 NBC 活动,而 CFTR 相对较低。此外,CFTR 的抑制导致主要酸碱转运蛋白和 CFTR 之间的功能相互作用。我们得出结论,EOs 为研究食管上皮细胞的离子转运机制提供了一个相关且合适的模型系统,并且它们也可以用作临床前工具来评估新的治疗化合物在与改变的离子转运过程相关的食管疾病中的有效性。
更新日期:2021-09-16
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