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Published on: February 12, 2019
Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis
So-Yi Chang1,2, Wen-Hsin Chang1,2, David C Yang1
1Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United States.
Abstract:
Precision-cut lung slices (PCLS) are commonly used as an ex vivo model to study lung fibrosis; however, traditional models lack immune cell infiltration, including the recruitment of monocytes and macrophages, which are critical for inflammation and fibrosis. To address this limitation, we developed novel autologous PCLS-immune co-culture models that better replicate the processes of inflammation, repair, and immune cell recruitment associated with fibrosis. Fibrotic responses to nicotine, cigarette smoke extract (CSE), and a fibrosis-inducing cocktail (FC) were first evaluated in PCLS containing only tissue-resident macrophages, with upregulation of α-SMA-expressing fibroblasts confirmed by immunofluorescence and Western blotting, and collagen deposition quantified using Sirius Red staining. To study macrophage recruitment, we employed an indirect co-culture model using transwells to approximate blood vessel function. Chemotactic studies revealed increased migration of autologous bone marrow-derived macrophages (BMDMs) toward and infiltration into CSE-injured PCLS. In a direct co-culture model simulating the repair phase of fibrosis, PCLS exposed to CSE and FC showed further increased collagen deposition in the presence of autologous BMDMs, but not heterologous ones. These findings suggest that our novel PCLS-immune co-culture models provide a platform for studying macrophage involvement in fibrosis and offer potential for developing macrophage-targeted therapeutic strategies in pulmonary fibrosis.
Insights
Researchers developed new co-culture models using precision-cut lung slices (PCLS) and immune cells to better study lung fibrosis. These models accurately show macrophage recruitment and collagen deposition, crucial for understanding fibrotic diseases.
Area of Science:
- Pulmonary Research
- Cell Biology
- Immunology
Background:
- Precision-cut lung slices (PCLS) are standard ex vivo models for lung fibrosis research.
- Traditional PCLS models lack immune cell infiltration, particularly monocytes and macrophages, which are key to fibrosis development.
Purpose of the Study:
- To develop novel autologous PCLS-immune co-culture models that mimic inflammation, repair, and immune cell recruitment in lung fibrosis.
- To investigate the role of macrophages in fibrotic responses induced by nicotine, cigarette smoke extract (CSE), and a fibrosis-inducing cocktail (FC).
Main Methods:
- Evaluated fibrotic responses in PCLS with tissue-resident macrophages using immunofluorescence, Western blotting, and Sirius Red staining.
- Utilized indirect co-culture transwell models to assess autologous bone marrow-derived macrophage (BMDM) migration and infiltration into CSE-injured PCLS.
- Employed direct co-culture models to examine collagen deposition in CSE/FC-exposed PCLS with autologous and heterologous BMDMs.
Main Results:
- Confirmed upregulation of α-SMA-expressing fibroblasts and increased collagen deposition in PCLS exposed to fibrotic stimuli.
- Demonstrated increased migration and infiltration of autologous BMDMs into CSE-injured PCLS via transwell assays.
- Observed enhanced collagen deposition in direct co-cultures of PCLS with autologous BMDMs, but not heterologous ones, during the repair phase.
Conclusions:
- The novel autologous PCLS-immune co-culture models effectively replicate key aspects of lung fibrosis, including immune cell recruitment and fibrotic matrix deposition.
- These models provide a valuable platform for studying macrophage involvement in pulmonary fibrosis.
- The findings suggest potential for developing macrophage-targeted therapies for lung fibrosis.
