Work

Protocol for the in vivo tracing of alveolar type I cells from mice using two distinct dual recombinase-mediated genetic approaches

Year: 2025

Type: article

Abstract: Revealing the origins of alveolar epithelial stem cells is crucial for preventing and treating lung diseases. Here, we present a protocol for tracing alveolar type I (AT1) cells from mice in vivo using two distinct dual recombinase-mediated systems. We describe steps for model establishment, harvesting tissue, cryosection, immunofluorescence staining, and confocal imaging. We then detail procedures for analysis of image files and quantification. This protocol provides a foundation for elucidating the plasticity of AT1 cell fate during homeostasis and diseases. For complete details on the use and execution of this protocol, please refer to Liu et al. 1 • Steps for characterizing the labeling profile of the single recombinase-based mouse lines • Procedure for developing dual recombinase-mediated nested mouse lines to label AT1 cells • Guidance on conducting fate mapping analysis of AT1 cells following lung injury Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Revealing the origins of alveolar epithelial stem cells is crucial for preventing and treating lung diseases. Here, we present a protocol for tracing alveolar type I (AT1) cells from mice in vivo using two distinct dual recombinase-mediated systems. We describe steps for model establishment, harvesting tissue, cryosection, immunofluorescence staining, and confocal imaging. We then detail procedures for analysis of image files and quantification. This protocol provides a foundation for elucidating the plasticity of AT1 cell fate during homeostasis and diseases.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.103648

Authors: Xueying Yang, Xinfeng Meng, Jingting Zhu, Zhongxiao Wang, Bin Zhou, Kuo Liu