Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis
Background: The use of BAL to study ILDs has improved our understanding of IPF pathogenesis. BAL fluid is routinely collected and can be considered a clinical and research tool. The procedure is well tolerated and minimally invasive. No specific cell lines from BAL or immortalized cell lines from IP...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-04-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/11/9/1441 |
| _version_ | 1827672912550690816 |
|---|---|
| author | Laura Bergantini Miriana d’Alessandro Sara Gangi Dalila Cavallaro Giuseppe Campiani Stefania Butini Claudia Landi Luca Bini Paolo Cameli Elena Bargagli |
| author_facet | Laura Bergantini Miriana d’Alessandro Sara Gangi Dalila Cavallaro Giuseppe Campiani Stefania Butini Claudia Landi Luca Bini Paolo Cameli Elena Bargagli |
| author_sort | Laura Bergantini |
| collection | DOAJ |
| description | Background: The use of BAL to study ILDs has improved our understanding of IPF pathogenesis. BAL fluid is routinely collected and can be considered a clinical and research tool. The procedure is well tolerated and minimally invasive. No specific cell lines from BAL or immortalized cell lines from IPF patients are available commercially. A method to quickly isolate and characterize fibroblasts from BAL is an unmet research need. Materials and methods: Here we describe a new protocol by which we isolated a cell line from IPF. The cell line was expanded in vitro and characterized phenotypically, morphologically and functionally. Results: This culture showed highly filamentous cells with an evident central nucleus. From the phenotypic point of view, this cell line displays fibroblast/myofibroblast-like features including expression of alpha-SMA, vimentin, collagen type-1 and fibronectin. The results showed high expression of ROS in these cells. Oxidative stress invariably promotes extracellular matrix expression in lung diseases directly or through over-production of pro-fibrotic growth factors. Conclusions: Our protocol makes it possible to obtain fibroblasts BAL that is a routine non-invasive method that offers the possibility of having a large sample of patients. Standardized culture methods are important for a reliable model for testing molecules and eventual novel development therapeutic targets. |
| first_indexed | 2024-03-10T04:16:56Z |
| format | Article |
| id | doaj.art-5f299860af564e97b2822137a9c8d64f |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T04:16:56Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-5f299860af564e97b2822137a9c8d64f2023-11-23T07:59:17ZengMDPI AGCells2073-44092022-04-01119144110.3390/cells11091441Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary FibrosisLaura Bergantini0Miriana d’Alessandro1Sara Gangi2Dalila Cavallaro3Giuseppe Campiani4Stefania Butini5Claudia Landi6Luca Bini7Paolo Cameli8Elena Bargagli9Respiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyRespiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyRespiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyRespiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyDepartment of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018–2022, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Biotechnology, Chemistry and Pharmacy, DoE Department of Excellence 2018–2022, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Life Sciences, University of Siena, 53100 Siena, ItalyDepartment of Life Sciences, University of Siena, 53100 Siena, ItalyRespiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyRespiratory Disease and Lung Transplant Unit, Department of Medical Sciences, Surgery and Neuroscience, Siena University, 53100 Siena, ItalyBackground: The use of BAL to study ILDs has improved our understanding of IPF pathogenesis. BAL fluid is routinely collected and can be considered a clinical and research tool. The procedure is well tolerated and minimally invasive. No specific cell lines from BAL or immortalized cell lines from IPF patients are available commercially. A method to quickly isolate and characterize fibroblasts from BAL is an unmet research need. Materials and methods: Here we describe a new protocol by which we isolated a cell line from IPF. The cell line was expanded in vitro and characterized phenotypically, morphologically and functionally. Results: This culture showed highly filamentous cells with an evident central nucleus. From the phenotypic point of view, this cell line displays fibroblast/myofibroblast-like features including expression of alpha-SMA, vimentin, collagen type-1 and fibronectin. The results showed high expression of ROS in these cells. Oxidative stress invariably promotes extracellular matrix expression in lung diseases directly or through over-production of pro-fibrotic growth factors. Conclusions: Our protocol makes it possible to obtain fibroblasts BAL that is a routine non-invasive method that offers the possibility of having a large sample of patients. Standardized culture methods are important for a reliable model for testing molecules and eventual novel development therapeutic targets.https://www.mdpi.com/2073-4409/11/9/1441BALfibroblastIPF |
| spellingShingle | Laura Bergantini Miriana d’Alessandro Sara Gangi Dalila Cavallaro Giuseppe Campiani Stefania Butini Claudia Landi Luca Bini Paolo Cameli Elena Bargagli Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis Cells BAL fibroblast IPF |
| title | Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis |
| title_full | Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis |
| title_fullStr | Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis |
| title_full_unstemmed | Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis |
| title_short | Bronchoalveolar-Lavage-Derived Fibroblast Cell Line (B-LSDM7) as a New Protocol for Investigating the Mechanisms of Idiopathic Pulmonary Fibrosis |
| title_sort | bronchoalveolar lavage derived fibroblast cell line b lsdm7 as a new protocol for investigating the mechanisms of idiopathic pulmonary fibrosis |
| topic | BAL fibroblast IPF |
| url | https://www.mdpi.com/2073-4409/11/9/1441 |
| work_keys_str_mv | AT laurabergantini bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT mirianadalessandro bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT saragangi bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT dalilacavallaro bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT giuseppecampiani bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT stefaniabutini bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT claudialandi bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT lucabini bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT paolocameli bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis AT elenabargagli bronchoalveolarlavagederivedfibroblastcelllineblsdm7asanewprotocolforinvestigatingthemechanismsofidiopathicpulmonaryfibrosis |