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从野生型小鼠中生成肺泡球体的优化方案。

An Optimized Protocol for the Generation of Alveolospheres from Wild-Type Mice.

机构信息

Department of Medicine V, Internal Medicine, Infectious Diseases and Infection Control, Universities of Giessen and Marburg Lung Center (UGMLC), German Center for Lung Research (DZL), Justus-Liebig University Giessen (JLU), 35392 Giessen, Germany.

Cardio-Pulmonary Institute (CPI), 35392 Giessen, Germany.

出版信息

Cells. 2024 May 27;13(11):922. doi: 10.3390/cells13110922.

Abstract

Organoid models have become an integral part of the research methodology in the lung field. These systems allow for the study of progenitor and stem cell self-renewal, self-organization, and differentiation. Distinct models of lung organoids mimicking various anatomical regions of mature lungs have emerged in parallel to the increased gain of knowledge regarding epithelial stem and progenitor cell populations and the corresponding mesenchymal cells that populate the in vivo niche. In the distal lung, type 2 alveolar epithelial cells (AEC2s) represent a stem cell population that is engaged in regenerative mechanisms in response to various insults. These cells self-renew and give rise to AEC1s that carry out gas exchange. Multiple experimental protocols allowing the generation of alveolar organoids, or alveolospheres, from murine lungs have been described. Among the drawbacks have been the requirement of transgenic mice allowing the isolation of AEC2s with high viability and purity, and the occasional emergence of bronchiolar and bronchioalveolar organoids. Here, we provide a refined gating strategy and an optimized protocol for the generation of alveolospheres from wild-type mice. Our approach not only overcomes the need for transgenic mice to generate such organoids, but also yields a pure culture of alveolospheres that is devoid of bronchiolar and bronchioalveolar organoids. Our protocol contributes to the standardization of this important research tool.

摘要

类器官模型已成为肺部研究方法学不可或缺的一部分。这些系统允许研究祖细胞和干细胞的自我更新、自我组织和分化。与对上皮干细胞和祖细胞群体以及在体内龛位中存在的相应间充质细胞的了解不断增加并行,出现了模拟成熟肺部不同解剖区域的不同肺类器官模型。在远端肺部,II 型肺泡上皮细胞 (AEC2) 代表了一个干细胞群体,该群体参与各种损伤的再生机制。这些细胞自我更新并产生执行气体交换的 AEC1。已经描述了多种允许从鼠肺中生成肺泡类器官或肺泡球体的实验方案。其中的缺点是需要能够分离具有高活力和纯度的 AEC2 的转基因小鼠,并且偶尔会出现细支气管和细支气管肺泡类器官。在这里,我们提供了一种从野生型小鼠生成肺泡球体的改良门控策略和优化方案。我们的方法不仅克服了生成此类类器官对转基因小鼠的需求,而且还产生了一种无细支气管和细支气管肺泡类器官的纯肺泡球体培养物。我们的方案有助于该重要研究工具的标准化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0130/11171706/9590f7305d56/cells-13-00922-g001.jpg

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