Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain
Abstract Introduction Fibrocytes are emerging myeloid‐derived circulating cells that can migrate into damaged tissues and usually contribute to their repair. Key features of fibrocytes include the expression myeloid markers, production of extracellular matrix proteins, and secretion of various humor...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-03-01
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| Series: | Immunity, Inflammation and Disease |
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| Online Access: | https://doi.org/10.1002/iid3.361 |
| _version_ | 1818573992307458048 |
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| author | Hiroshi Kawano Kazuya Koyama Haruka Nishimura Yuko Toyoda Kozo Kagawa Seidai Sato Nobuhito Naito Hisatsugu Goto Yutaka Inagaki Yasuhiko Nishioka |
| author_facet | Hiroshi Kawano Kazuya Koyama Haruka Nishimura Yuko Toyoda Kozo Kagawa Seidai Sato Nobuhito Naito Hisatsugu Goto Yutaka Inagaki Yasuhiko Nishioka |
| author_sort | Hiroshi Kawano |
| collection | DOAJ |
| description | Abstract Introduction Fibrocytes are emerging myeloid‐derived circulating cells that can migrate into damaged tissues and usually contribute to their repair. Key features of fibrocytes include the expression myeloid markers, production of extracellular matrix proteins, and secretion of various humoral factors that activate resident fibroblasts; they also have the potential to differentiate into fibroblasts. However, no specific surface markers have been identified to identify fibrocytes in vivo. One reason could be that the method used to detect fibrocytes requires intracellular collagen staining. Methods In the present study, to establish an improved method for the detection of lung fibrocytes and to analyze viable fibrocytes, we used collagen I(α)2‐green fluorescent protein (Col‐GFP) reporter mice, which had undergone the intratracheal instillation of bleomycin (BLM). Results Using flow cytometry to gate out cells with autofluorescence, we clearly found that CD45+ GFP+ cells resided in the lungs of Col‐GFP mice at a steady state and these cells increased after BLM injury, peaking at Day 14. These cells expressed not only known cell surface markers of fibrocytes, but also some novel markers, in addition to a low level of collagen I in comparison to CD45− GFP+ cells. Conclusion Our findings suggest that the improved method can be a useful for the detection of pure lung fibrocytes and allows us to further analyze the characteristics of viable fibrocytes. |
| first_indexed | 2024-12-15T00:20:29Z |
| format | Article |
| id | doaj.art-cdd4007fc96349ecb3ca78bae2e67825 |
| institution | Directory Open Access Journal |
| issn | 2050-4527 |
| language | English |
| last_indexed | 2024-12-15T00:20:29Z |
| publishDate | 2021-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Immunity, Inflammation and Disease |
| spelling | doaj.art-cdd4007fc96349ecb3ca78bae2e678252022-12-21T22:42:20ZengWileyImmunity, Inflammation and Disease2050-45272021-03-019112012710.1002/iid3.361Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strainHiroshi Kawano0Kazuya Koyama1Haruka Nishimura2Yuko Toyoda3Kozo Kagawa4Seidai Sato5Nobuhito Naito6Hisatsugu Goto7Yutaka Inagaki8Yasuhiko Nishioka9Department of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanDepartment of Regenerative Medicine, Center for Matrix Biology and Medicine, Graduate School of Medicine Tokai University Isehara JapanDepartment of Respiratory Medicine & Rheumatology, Graduate School of Biomedical Sciences Tokushima University Tokushima JapanAbstract Introduction Fibrocytes are emerging myeloid‐derived circulating cells that can migrate into damaged tissues and usually contribute to their repair. Key features of fibrocytes include the expression myeloid markers, production of extracellular matrix proteins, and secretion of various humoral factors that activate resident fibroblasts; they also have the potential to differentiate into fibroblasts. However, no specific surface markers have been identified to identify fibrocytes in vivo. One reason could be that the method used to detect fibrocytes requires intracellular collagen staining. Methods In the present study, to establish an improved method for the detection of lung fibrocytes and to analyze viable fibrocytes, we used collagen I(α)2‐green fluorescent protein (Col‐GFP) reporter mice, which had undergone the intratracheal instillation of bleomycin (BLM). Results Using flow cytometry to gate out cells with autofluorescence, we clearly found that CD45+ GFP+ cells resided in the lungs of Col‐GFP mice at a steady state and these cells increased after BLM injury, peaking at Day 14. These cells expressed not only known cell surface markers of fibrocytes, but also some novel markers, in addition to a low level of collagen I in comparison to CD45− GFP+ cells. Conclusion Our findings suggest that the improved method can be a useful for the detection of pure lung fibrocytes and allows us to further analyze the characteristics of viable fibrocytes.https://doi.org/10.1002/iid3.361CD11bCD11cfibrocyteGr‐1pulmonary fibrosis |
| spellingShingle | Hiroshi Kawano Kazuya Koyama Haruka Nishimura Yuko Toyoda Kozo Kagawa Seidai Sato Nobuhito Naito Hisatsugu Goto Yutaka Inagaki Yasuhiko Nishioka Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain Immunity, Inflammation and Disease CD11b CD11c fibrocyte Gr‐1 pulmonary fibrosis |
| title | Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| title_full | Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| title_fullStr | Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| title_full_unstemmed | Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| title_short | Development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| title_sort | development of improved method to identify and analyze lung fibrocytes with flow cytometry in a reporter mouse strain |
| topic | CD11b CD11c fibrocyte Gr‐1 pulmonary fibrosis |
| url | https://doi.org/10.1002/iid3.361 |
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