Simultaneous isolation of proximal and distal lung progenitor cells from individual mice using a 3D printed guide reduces proximal cell contamination of distal lung epithelial cell isolations.

Autor:	Alsafadi HN; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden., Stegmayr J; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden., Ptasinski V; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden; Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden., Silva I; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden., Mittendorfer M; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden., Murray LA; Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden; Research and Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Cambridge, UK., Wagner DE; Department of Experimental Medical Sciences, Faculty of Medicine, Lund University, Lund, Sweden; Wallenberg Centre for Molecular Medicine, Faculty of Medicine, Lund University, Lund, Sweden; Stem Cell Center, Faculty of Medicine, Lund University, Lund, Sweden; NanoLund, Lund University, Lund, Sweden. Electronic address: darcy.wagner@med.lu.se.
Jazyk:	angličtina
Zdroj:	Stem cell reports [Stem Cell Reports] 2022 Dec 13; Vol. 17 (12), pp. 2718-2731. Date of Electronic Publication: 2022 Dec 01.
DOI:	10.1016/j.stemcr.2022.11.002
Abstrakt:	The respiratory epithelium consists of multiple, functionally distinct cell types and is maintained by regionally specific progenitor populations that repair the epithelium following injury. Several in vitro methods exist for studying lung epithelial repair using primary murine lung cells, but isolation methods are hampered by a lack of surface markers distinguishing epithelial progenitors along the respiratory epithelium. Here, we developed a 3D printed lobe divider (3DLD) to aid in simultaneous isolation of proximal versus distal lung epithelial progenitors from individual mice that give rise to differentiated epithelia in multiple in vitro assays. In contrast to 3DLD-isolated distal progenitor cells, commonly used manual tracheal ligation methods followed by lobe removal resulted in co-isolation of rare proximal cells with distal cells, which altered the transcriptional landscape and size distribution of distal organoids. The 3DLD aids in reproducible isolation of distal versus proximal progenitor populations and minimizes the potential for contaminating populations to confound in vitro assays. Competing Interests: Conflict of interests V.P. is a full-time employee and received research funding at AstraZeneca and is a shareholder in AstraZeneca. At the time of contribution, L.M. was a full-time employee and received research funding at AstraZeneca. (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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