Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis
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...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphys.2025.1526787/full |
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| author | So-Yi Chang So-Yi Chang Wen-Hsin Chang Wen-Hsin Chang David C. Yang Qi-Sheng Hong Qi-Sheng Hong Ssu-Wei Hsu Ssu-Wei Hsu Reen Wu Ching-Hsien Chen Ching-Hsien Chen |
| author_facet | So-Yi Chang So-Yi Chang Wen-Hsin Chang Wen-Hsin Chang David C. Yang Qi-Sheng Hong Qi-Sheng Hong Ssu-Wei Hsu Ssu-Wei Hsu Reen Wu Ching-Hsien Chen Ching-Hsien Chen |
| author_sort | So-Yi Chang |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-76d39e394aec4c06b424a101f58032ed |
| institution | Kabale University |
| issn | 1664-042X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj-art-76d39e394aec4c06b424a101f58032ed2025-01-31T06:39:58ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2025-01-011610.3389/fphys.2025.15267871526787Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosisSo-Yi Chang0So-Yi Chang1Wen-Hsin Chang2Wen-Hsin Chang3David C. Yang4Qi-Sheng Hong5Qi-Sheng Hong6Ssu-Wei Hsu7Ssu-Wei Hsu8Reen Wu9Ching-Hsien Chen10Ching-Hsien Chen11Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Nephrology, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Nephrology, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Nephrology, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Nephrology, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesDivision of Nephrology, Department of Internal Medicine, University of California Davis, Davis, CA, United StatesPrecision-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.https://www.frontiersin.org/articles/10.3389/fphys.2025.1526787/fullprecision-cut lung slices (PCLS)macrophage recruitmentfibrosis inducersco-culture modelspulmonary fibrosis |
| spellingShingle | So-Yi Chang So-Yi Chang Wen-Hsin Chang Wen-Hsin Chang David C. Yang Qi-Sheng Hong Qi-Sheng Hong Ssu-Wei Hsu Ssu-Wei Hsu Reen Wu Ching-Hsien Chen Ching-Hsien Chen Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis Frontiers in Physiology precision-cut lung slices (PCLS) macrophage recruitment fibrosis inducers co-culture models pulmonary fibrosis |
| title | Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis |
| title_full | Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis |
| title_fullStr | Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis |
| title_full_unstemmed | Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis |
| title_short | Autologous precision-cut lung slice co-culture models for studying macrophage-driven fibrosis |
| title_sort | autologous precision cut lung slice co culture models for studying macrophage driven fibrosis |
| topic | precision-cut lung slices (PCLS) macrophage recruitment fibrosis inducers co-culture models pulmonary fibrosis |
| url | https://www.frontiersin.org/articles/10.3389/fphys.2025.1526787/full |
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