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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

Alsafadi, Hani N LU orcid ; Stegmayr, John LU ; Ptasinski, Victoria LU ; Silva, Iran LU orcid ; Mittendorfer, Margareta LU ; Murray, Lynne A and Wagner, Darcy E LU orcid (2022) In Stem Cell Reports 17(12). p.2718-2731
Abstract

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... (More)

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.

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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Stem Cell Reports
volume
17
issue
12
pages
2718 - 2731
publisher
Cell Press
external identifiers
  • pmid:36460000
  • scopus:85143875009
ISSN
2213-6711
DOI
10.1016/j.stemcr.2022.11.002
language
English
LU publication?
yes
additional info
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
id
680b88cc-0d31-40be-ab19-db90aa2688c5
date added to LUP
2022-12-04 20:12:47
date last changed
2024-04-04 13:26:00
@article{680b88cc-0d31-40be-ab19-db90aa2688c5,
  abstract     = {{<p>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.</p>}},
  author       = {{Alsafadi, Hani N and Stegmayr, John and Ptasinski, Victoria and Silva, Iran and Mittendorfer, Margareta and Murray, Lynne A and Wagner, Darcy E}},
  issn         = {{2213-6711}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{12}},
  pages        = {{2718--2731}},
  publisher    = {{Cell Press}},
  series       = {{Stem Cell Reports}},
  title        = {{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}},
  url          = {{http://dx.doi.org/10.1016/j.stemcr.2022.11.002}},
  doi          = {{10.1016/j.stemcr.2022.11.002}},
  volume       = {{17}},
  year         = {{2022}},
}