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Research: Cell culture system to assay candidate genes and molecular pathways implicated in congenital diaphragmatic hernias

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Research: Cell culture system to assay candidate genes and molecular pathways implicated in congenital diaphragmatic hernias

Dev Biol

. 2020 Aug 19;S0012-1606(20)30209-8. doi: 10.1016/j.ydbio.2020.07.013. Online ahead of print https://pubmed.ncbi.nlm.nih.gov/32827499/.

Cell culture system to assay candidate genes and molecular pathways implicated in congenital diaphragmatic hernias

Eric L Bogenschutz 1Elizabeth M Sefton 1Gabrielle Kardon 2Affiliations expand

Abstract

The mammalian muscularized diaphragm is essential for respiration and defects in the developing diaphragm cause a common and frequently lethal birth defect, congenital diaphragmatic hernia (CDH). Human genetic studies have implicated more than 150 genes and multiple molecular pathways in CDH, but few of these have been validated because of the expense and time to generate mouse mutants. The pleuroperitoneal folds (PPFs) are transient embryonic structures in diaphragm development and defects in PPFs lead to CDH. We have developed a system to culture PPF fibroblasts from E12.5 mouse embryos and show that these fibroblasts, in contrast to the commonly used NIH 3T3 fibroblasts, maintain expression of key genes in normal diaphragm development. Using pharmacological and genetic manipulations that result in CDH in vivo, we also demonstrate that differences in proliferation provide a rapid means of distinguishing healthy and impaired PPF fibroblasts. Thus, the PPF fibroblast cell culture system is an efficient tool for assaying the functional significance of CDH candidate genes and molecular pathways and will be an important resource for elucidating the complex etiology of CDH.

Keywords: CDH; Cell culture; Congenital diaphragmatic hernia; Diaphragm; Fibroblasts.

Copyright © 2020. Published by Elsevier Inc.

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