Journal article
Loss of lung mesenchymal identities in culture and a screening approach to identify restorative cues
American journal of physiology. Lung cellular and molecular physiology, Vol.329(6), pp.L732-L748
12/2025
DOI: 10.1152/ajplung.00177.2025
PMCID: PMC12754774
PMID: 41213154
Abstract
Lung fibroblasts generate and respond to mechanical, biochemical, and matrix cues present in their microenvironment. With the advent of next-generation sequencing technologies, multiple studies describe transcriptionally unique fibroblast subpopulations in the human lung. However, limited published data suggest a loss of fibroblast native phenotypes and functions after culture ex vivo. In this study, we characterized changes in transcriptional programs of human lung mesenchyme isolated from freshly procured tissue and maintained in traditional cell culture conditions. Our results demonstrate that fibroblasts isolated and cultured in this manner adopt transcriptional programs largely distinct from those observed
. To recapitulate distinct native fibroblast states
, we sought to develop a screening approach to identify cues promoting native fibroblast identities. From published single-cell data, we identified unique transcriptional markers of alveolar and adventitial fibroblast subpopulations and validated the sensitivity of ELISAs for detecting changes in secreted markers of these fibroblast subpopulations. We then stimulated primary human lung fibroblasts with soluble cues known to act on fibroblasts, quantifying changes in secreted and transcriptional markers by ELISA and qPCR. While our small pilot screen did not identify single cues capable of fully recapitulating fibroblast
states, it established a system that can be expanded to broadly screen additional cues and pointed toward factors likely to be critical in developing better culture models for studying human lung fibroblast function and plasticity.
Details
- Title: Subtitle
- Loss of lung mesenchymal identities in culture and a screening approach to identify restorative cues
- Creators
- Ana M Diaz Espinosa - Mayo ClinicJack H Wellmerling - Mayo ClinicNatalia Del Pilar Vanegas - The Ohio State UniversityJhonny Rodriguez-Lopez - The Ohio State UniversityKyoung M Choi - Mayo ClinicNichole Aquna Schussler - Mayo ClinicPatrick A Link - Mayo ClinicLuis R Rodriguez - University of PennsylvaniaGrant Barringer - Mayo ClinicMauricio Rojas - The Ohio State UniversityDaniel J Tschumperlin - Mayo Clinic
- Resource Type
- Journal article
- Publication Details
- American journal of physiology. Lung cellular and molecular physiology, Vol.329(6), pp.L732-L748
- DOI
- 10.1152/ajplung.00177.2025
- PMID
- 41213154
- PMCID
- PMC12754774
- NLM abbreviation
- Am J Physiol Lung Cell Mol Physiol
- ISSN
- 1522-1504
- eISSN
- 1522-1504
- Publisher
- American Physiological Society
- Grant note
- NIH HL166187 / HHS | NIH | National Heart, Lung, and Blood Institute (NHLBI) NIH HL152967 / HHS | NIH | National Heart, Lung, and Blood Institute (NHLBI) NIH K99 HL171946 / HHS | NIH | National Heart, Lung, and Blood Institute (NHLBI)
- Language
- English
- Electronic publication date
- 11/10/2025
- Date published
- 12/2025
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Craniofacial Anomalies Research Center
- Record Identifier
- 9985026466202771
Metrics
16 Record Views