Gregory Fleming James Cystic Fibrosis Research Center, Univ. of Alabama at Birmingham, Birmingham, AL, United States of America.
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, United States of America.
PLoS One. 2023 Oct 24;18(10):e0293367. doi: 10.1371/journal.pone.0293367. eCollection 2023.
Cystic fibrosis (CF) is a genetic disease hallmarked by aberrant ion transport that results in delayed mucus clearance, chronic infection, and progressive lung function decline. Several animal models have been developed to study the airway anatomy and mucus physiology in CF, but they are costly and difficult to maintain, making them less accessible for many applications. A more available CFTR-/- rat model has been developed and characterized to develop CF airway abnormalities, but consistent dosing of pharmacologic agents and longitudinal evaluation remain a challenge. In this study, we report the development and characterization of a novel ex vivo trachea model that utilizes both wild type (WT) and CFTR-/- rat tracheae cultured on a porcine gelatin matrix. Here we show that the ex vivo tracheae remain viable for weeks, maintain a CF disease phenotype that can be readily quantified, and respond to stimulation of mucus and fluid secretion by cholinergic stimulation. Furthermore, we show that ex vivo tracheae may be used for well-controlled pharmacological treatments, which are difficult to perform on freshly excised trachea or in vivo models with this degree of scrutiny. With improved interrogation possible with a durable trachea, we also established firm evidence of a gland secretion defect in CFTR-/- rat tracheae compared to WT controls. Finally, we demonstrate that the ex vivo tracheae can be used to generate high mucus protein yields for subsequent studies, which are currently limited by in vivo mucus collection techniques. Overall, this study suggests that the ex vivo trachea model is an effective, easy to set up culture model to study airway and mucus physiology.
囊性纤维化（CF）是一种遗传性疾病，其特征是离子转运异常，导致黏液清除延迟、慢性感染和肺功能进行性下降。已经开发了几种动物模型来研究 CF 的气道解剖结构和黏液生理学，但它们成本高且难以维持，因此许多应用都难以获得。已经开发并表征了一种更易于获得的 CFTR-/-大鼠模型，以开发 CF 气道异常，但药物的一致给药和纵向评估仍然是一个挑战。在这项研究中，我们报告了一种新型离体气管模型的开发和表征，该模型利用在猪明胶基质上培养的野生型（WT）和 CFTR-/-大鼠气管。在这里，我们表明离体气管可以存活数周，保持 CF 疾病表型，并且可以很容易地进行定量，并且对胆碱能刺激引起的黏液和液体分泌的刺激有反应。此外，我们表明离体气管可用于进行良好控制的药理学治疗，而这在新鲜取出的气管或具有这种严格程度的体内模型中很难进行。随着对持久气管的更好的询问，我们还为 CFTR-/-大鼠气管与 WT 对照相比的腺体分泌缺陷提供了确凿的证据。最后，我们证明离体气管可用于产生高黏液蛋白产量，以进行随后的研究，这目前受到体内黏液收集技术的限制。总体而言，这项研究表明，离体气管模型是一种有效的、易于建立的培养模型，可用于研究气道和黏液生理学。
