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

Bogenschutz, Eric L.; Sefton, Elizabeth M.; Kardon, Gabrielle

doi:10.1016/j.ydbio.2020.07.013

Cited by 8 publications

(10 citation statements)

References 54 publications

(61 reference statements)

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“…CDH cannot be attributed to one single source or defect, which makes it hard to study its pathogenesis, but new cell culture models can aid in improving insights at the cellular level. Recently, a cell culture model was described where PPFs from mice were isolated and cultured to outgrow and expand PPF fibroblasts, that maintained expression of key diaphragm genes ( 43 ). Pharmacological inhibition or genetic manipulation that causes CDH resulted in reduced in vitro proliferation in PPF-derived fibroblasts ( 43 ).…”

Section: Cellular Models In Cdh Researchmentioning

confidence: 99%

Cellular Origin(s) of Congenital Diaphragmatic Hernia

et al. 2021

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Congenital diaphragmatic hernia (CDH) is a structural birth defect characterized by a diaphragmatic defect, lung hypoplasia and structural vascular defects. In spite of recent developments, the pathogenesis of CDH is still poorly understood. CDH is a complex congenital disorder with multifactorial etiology consisting of genetic, cellular and mechanical factors. This review explores the cellular origin of CDH pathogenesis in the diaphragm and lungs and describes recent developments in basic and translational CDH research.

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Section: Cellular Models In Cdh Researchmentioning

confidence: 99%

Cellular Origin(s) of Congenital Diaphragmatic Hernia

et al. 2021

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“…To investigate the function of Met in diaphragm muscle progenitors, we turned to an in vitro system to co-culture E12.5 diaphragm fibroblasts and myoblasts (Bogenschutz et al, 2020) in combination with BMS777607. PPFs were dissected from E12.5 Pax3 CreKI/+ ; Rosa nTnG/+ embryos (Engleka et al, 2005;Prigge et al, 2013), in which Pax3-derived myogenic nuclei are GFP+ and PPF fibroblast nuclei are Tomato+, and cultured with either 10 μM BMS777607 or DMSO vehicle control.…”

Section: Met Signaling Is Required For Survival Motility and Commitment Of Diaphragm Muscle Progenitors In Vitromentioning

confidence: 99%

“…Cell culture, media and reagents E12.5 embryos were dissected from pregnant Rosa mTmG/mTmG or Rosa nTnG/nTnG females mated with Pax3 CreKI/+ males. Embryos and PPFs were dissected as previously described (Bogenschutz et al, 2020). Growth of both PPF fibroblasts and myogenic cells was promoted using DMEM/F-12 GlutaMAX (Invitrogen), 10% FBS, 50 μg/mL gentamycin, and 0.5nM FGF.…”

Section: Microscopy and Three-dimensional Renderingmentioning

confidence: 99%

“…Using conditional mutagenesis, pharmacological treatments, and an in vitro primary cell culture system (Bogenschutz et al, 2020), we demonstrate that the diaphragm's connective tissue fibroblasts are a critical source of HGF that recruits and maintains muscle progenitors into and throughout the developing diaphragm and controls defasciculation of the phrenic nerve.…”

Section: Introductionmentioning

confidence: 99%

“…Using conditional mutagenesis, pharmacological treatments, and an in vitro primary cell culture system (Bogenschutz et al, 2020), we demonstrate that the diaphragm's (which was not certified by peer review) is the author/funder. All rights reserved.…”

Section: Introductionmentioning

confidence: 99%

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Fibroblast-derived HGF integrates muscle and nerve development during morphogenesis of the mammalian diaphragm

Sefton

Gallardo

et al. 2021

Preprint

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The diaphragm is a domed muscle between the thorax and abdomen essential for breathing in mammals. Diaphragm development requires the coordinated development of muscle, connective tissue, and nerve, which are derived from different embryonic sources. Defects in diaphragm development cause the common and often lethal birth defect, Congenital Diaphragmatic Hernias (CDH). HGF/MET signaling is required for diaphragm muscularization, but the source of HGF and the specific functions of this pathway in muscle progenitors or potentially the phrenic nerve have not been explicitly tested. Using conditional mutagenesis and pharmacological inhibition of MET, we demonstrate that the pleuroperitoneal folds (PPFs), transient embryonic structures that give rise to the connective tissue, are the source of HGF critical for diaphragm muscularization and phrenic nerve primary branching. HGF not only is required for recruitment of muscle progenitors to the diaphragm, but is continuously required for maintenance and motility of the pool of progenitors to enable full muscularization. Thus, the connective tissue fibroblasts and HGF coordinately regulate diaphragm muscularization and innervation. Defects in PPF-derived HGF result in muscleless regions that are susceptible to CDH.

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The role of genes and environment in the etiology of congenital diaphragmatic hernias

Burns¹,

Kardon²

2023

Current Topics in Developmental Biology

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

Abstract: Multiple candidate genes and molecular pathways have been identified as potentially contributing to the etiology of congenital diaphragmatic hernias. We describe a cell culture system to efficiently functionally test these candidates.

Cited by 8 publications

References 54 publications

Cellular Origin(s) of Congenital Diaphragmatic Hernia

Cellular Origin(s) of Congenital Diaphragmatic Hernia

Fibroblast-derived HGF integrates muscle and nerve development during morphogenesis of the mammalian diaphragm

The role of genes and environment in the etiology of congenital diaphragmatic hernias

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