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Selective activation and proliferation of a quiescent stem cell population in the neuroepithelial body microenvironment.

Respiratory research (2018-10-28)
Line Verckist, Isabel Pintelon, Jean-Pierre Timmermans, Inge Brouns, Dirk Adriaensen
ABSTRACT

The microenvironment (ME) of neuroepithelial bodies (NEBs) harbors densely innervated groups of pulmonary neuroendocrine cells that are covered by Clara-like cells (CLCs) and is believed to be important during development and for adult airway epithelial repair after severe injury. Yet, little is known about its potential stem cell characteristics in healthy postnatal lungs. Transient mild lung inflammation was induced in mice via a single low-dose intratracheal instillation of lipopolysaccharide (LPS). Bronchoalveolar lavage fluid (BALF), collected 16 h after LPS instillation, was used to challenge the NEB ME in ex vivo lung slices of control mice. Proliferating cells in the NEB ME were identified and quantified following simultaneous LPS instillation and BrdU injection. The applied LPS protocol induced very mild and transient lung injury. Challenge of lung slices with BALF of LPS-treated mice resulted in selective Ca2+-mediated activation of CLCs in the NEB ME of control mice. Forty-eight hours after LPS challenge, a remarkably selective and significant increase in the number of divided (BrdU-labeled) cells surrounding NEBs was observed in lung sections of LPS-challenged mice. Proliferating cells were identified as CLCs. A highly reproducible and minimally invasive lung inflammation model was validated for inducing selective activation of a quiescent stem cell population in the NEB ME. The model creates new opportunities for unraveling the cellular mechanisms/pathways regulating silencing, activation, proliferation and differentiation of this unique postnatal airway epithelial stem cell population.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
5-Bromo-2′-deoxyuridine, ≥99% (HPLC)
Sigma-Aldrich
Lipopolysaccharides from Escherichia coli O55:B5, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%
Sigma-Aldrich
Agarose, low gelling temperature
Sigma-Aldrich
4-(4-Diethylaminostyryl)-1-methylpyridinium iodide, ≥97%

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