Culture Conditions Differentially Regulate the Inflammatory Niche and Cellular Phenotype of Tracheo-Bronchial Basal Stem Cells DOI Creative Commons

Shubha Murthy,

Denise A Seabold,

Lalit Kumar Gautam

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 5, 2024

ABSTRACT Human bronchial epithelial cells (HBECs) derived from the tracheo-bronchial regions of human airways provide an excellent in vitro model for studying pathological mechanisms and evaluating therapeutics airway cells. This cell population comprises a mixed basal (BCs), predominant stem capable both self-renewal functional differentiation. Despite their potential regenerative medicine, BCs exhibit significant phenotypic variability culture. To investigate how culture conditions influence BC phenotype function, we expanded three independent isolates media, growth medium (AECGM), dual-SMAD inhibitor (DSI)-enriched AECGM, Pneumacult Ex plus (PEx+). Extensive RNA sequencing, immune assays electrical measurements revealed that PEx+ media significantly drove proliferation broad pro-inflammatory BCs. In contrast, displayed increased expression structural extracellular matrix components at high passage. Whereas AECGM some cytokines passage, DSI suppressed inflammation altogether thus implicating TGF-β inflammatory processes. Differentiation capacity declined with time irrespective expansion except PLUNC expressing secretory were elevated passage suggestive modulatory role These findings underscore profound impact on niche function The driven by particular, should be considered development cell-based models diseases therapeutic application. NEW & NOTEWORTHY Airway cells, vital regeneration therapies, show changes based conditions. Our study reveals composition duration greatly affect properties deposition results critical phenotype, influencing disease research therapy.

Language: Английский

High-throughput Mucus Microrheology for Phenotyping and Disease Modeling DOI Creative Commons
Feng Ling, Ayse T. Sahin, Bernardo Miller Naranjo

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 14, 2025

Mucus plays an integral role for the barrier function of many epithelial tissues. In human airways, mucus is constantly secreted to capture inhaled microbes and pollutants cleared away through concerted ciliary motion. Many important respiratory diseases exhibit altered flowability impaired clearance, contributing distress increased risk infections. Understanding how rheology changes during disease progression in response treatments thus great interest subtyping patients tailoring treatments, probing mechanisms, therapies; however, basic research greatly hampered by lack scalable user-friendly rheometry assays small volumes typically produced vitro models clinical ex vivo settings. To address this challenge, we developed a streamlined, high-throughput protocol leveraging Differential Dynamic Microscopy (DDM) reliably measure frequency-dependent microrheology minuscule (3-10 µ L) samples using standard epifluorescence microscopy. Our method does not require time-consuming user-interventions common particle tracking routines measures at time scale relaxation (1-20s), hence reducing assay time. We demonstrate successful application our harvested from state-of-art air-liquid-interface (ALI) cultures assess airway different culture conditions. show that approach equally applies other types sources mucus, also validated cervical mucus. envision can be seamlessly adopted non-expert users, without need specialized equipment or extensive training, study their respiratory, intestinal, reproductive mucosal organ systems. This advancement opens up new avenues large-scale studies, providing insights into previously limited data accessibility resource constraints.

Language: Английский

Citations

0
Physiology and pathophysiology of mucus and mucolytic use in critically ill patients DOI Creative Commons
Thomas Roe, Thomas R. Talbot,

Isis Terrington

et al.

Critical Care, Journal Year: 2025, Volume and Issue: 29(1)

Published: Feb. 7, 2025

Abstract Airway mucus is a highly specialised secretory fluid which functions as physical and immunological barrier to pathogens whilst lubricating the airways humifying atmospheric air. Dysfunction common during critical illness characterised by changes in production rate, chemical composition, properties, inflammatory phenotype. Mucociliary clearance, determined part characteristics ciliary function, also dysfunctional via disease related iatrogenic mechanisms. The consequences of dysfunction are potentially devastating, contributing prolonged ventilator dependency, increased risk secondary pneumonia, worsened lung injury. Mucolytic therapies designed decrease viscosity, improve expectoration/suctioning, thereby promote removal. Mucolytics, including hypertonic saline, dornase alfa/rhDNase, nebulised heparin, carbocisteine/N-Acetyl cysteine, commonly used critically ill patients. This review summarises physiology pathophysiology existing evidence for use mucolytics patients speculates on journey individualised mucolytic therapy.

Language: Английский

Citations

0
Characterization of a primary cellular airway model for inhalative drug delivery in comparison with the established permanent cell lines CaLu3 and RPMI 2650 DOI Creative Commons
Janik Martin,

Rebecca Rittersberger,

Simon Treitler

et al.

In vitro models, Journal Year: 2024, Volume and Issue: 3(4-6), P. 183 - 203

Published: Nov. 25, 2024

Language: Английский

Citations

1
Culture Conditions Differentially Regulate the Inflammatory Niche and Cellular Phenotype of Tracheo-Bronchial Basal Stem Cells DOI Creative Commons

Shubha Murthy,

Denise A Seabold,

Lalit Kumar Gautam

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 5, 2024

ABSTRACT Human bronchial epithelial cells (HBECs) derived from the tracheo-bronchial regions of human airways provide an excellent in vitro model for studying pathological mechanisms and evaluating therapeutics airway cells. This cell population comprises a mixed basal (BCs), predominant stem capable both self-renewal functional differentiation. Despite their potential regenerative medicine, BCs exhibit significant phenotypic variability culture. To investigate how culture conditions influence BC phenotype function, we expanded three independent isolates media, growth medium (AECGM), dual-SMAD inhibitor (DSI)-enriched AECGM, Pneumacult Ex plus (PEx+). Extensive RNA sequencing, immune assays electrical measurements revealed that PEx+ media significantly drove proliferation broad pro-inflammatory BCs. In contrast, displayed increased expression structural extracellular matrix components at high passage. Whereas AECGM some cytokines passage, DSI suppressed inflammation altogether thus implicating TGF-β inflammatory processes. Differentiation capacity declined with time irrespective expansion except PLUNC expressing secretory were elevated passage suggestive modulatory role These findings underscore profound impact on niche function The driven by particular, should be considered development cell-based models diseases therapeutic application. NEW & NOTEWORTHY Airway cells, vital regeneration therapies, show changes based conditions. Our study reveals composition duration greatly affect properties deposition results critical phenotype, influencing disease research therapy.

Language: Английский

Citations

0