Establishment and Characterization of a Novel Fibroblastic Cell Line (SCI13D) Derived from the Broncho-Alveolar Lavage of a Patient with Fibrotic Hypersensitivity Pneumonitis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Patient, Bronchoalveolar Fluid Collection, and Histological Sample Assessment
2.2. Cell Line Establishment and Culture
2.3. mRNA Extraction and Quantitative Real-Time RT-PCR
2.4. Morphology, Cyto-Fluorographic, and Immunofluorescence Analyses
2.5. Immunocytochemistry of the SCI13D Cell Line
2.6. Proliferation and Growth Kinetic Determination
2.7. Wound Sratch Test
2.8. Determination of the Total Cellular F-Actin with/without Pirfenidone Treatment
2.9. β-Galactosidase Assessment
2.10. Cytogenetics
2.11. Short Tandem Repeat Analysis
2.12. Statistical Analysis
3. Results
3.1. Patient’s Clinical and Diagnostic Features
3.2. Establishment of a Unique Fibroblastic Cell Line (SCI13D) from fHP Patient
3.3. Immunocytochemical Characteristic of the SCI13D Cell Line
3.4. Karyotype and FISH
3.5. Authenticity of the SCI13D Cell Line
3.6. Inhibition of Proliferation by Antifibrotic Therapy
3.7. Pirfenidone Slightly Inhibited Type 1 Collagen Expression in TGF-β-Treated SCI13D Cells
3.8. Inhibition of SCI13D Migration by Pirfenidone
3.9. Reduction of the TGFβ-Induced Profibrotic Phenotype by Pirfenidone in SCI13D Cell Line
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Cells Assessed | ||
---|---|---|
STR Locus | PBL | SCI13D |
D5S818 | 11,12 | 11,12 |
D13S17 | 9,11 | 9,11 |
D7S820 | 11 | 11 |
D16S539 | 9,14 | 9,14 |
VWA | 16 | 16 |
TH01 | 6 | 6 |
AM | x, y | x, y |
TPOX | 9,10 | 9,10 |
CSF1PO | 11,12 | 11,12 |
D21S11 | 27,30 | 27,30 |
D3S1358 | 16 | 16 |
D18S51 | 13,16 | 13,16 |
Penta E | 7,17 | 7,17 |
Penta D | 9,11 | 9,11 |
D8S1179 | 14,15 | 14,15 |
FGA | 19,22 | 19,22 |
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Giannoni, P.; Grosso, M.; Fugazza, G.; Nizzari, M.; Capra, M.C.; Bianchi, R.; Fiocca, R.; Salvi, S.; Montecucco, F.; Bertolotto, M.; et al. Establishment and Characterization of a Novel Fibroblastic Cell Line (SCI13D) Derived from the Broncho-Alveolar Lavage of a Patient with Fibrotic Hypersensitivity Pneumonitis. Biomedicines 2021, 9, 1193. https://doi.org/10.3390/biomedicines9091193
Giannoni P, Grosso M, Fugazza G, Nizzari M, Capra MC, Bianchi R, Fiocca R, Salvi S, Montecucco F, Bertolotto M, et al. Establishment and Characterization of a Novel Fibroblastic Cell Line (SCI13D) Derived from the Broncho-Alveolar Lavage of a Patient with Fibrotic Hypersensitivity Pneumonitis. Biomedicines. 2021; 9(9):1193. https://doi.org/10.3390/biomedicines9091193
Chicago/Turabian StyleGiannoni, Paolo, Marco Grosso, Giuseppina Fugazza, Mario Nizzari, Maria Cristina Capra, Rita Bianchi, Roberto Fiocca, Sandra Salvi, Fabrizio Montecucco, Maria Bertolotto, and et al. 2021. "Establishment and Characterization of a Novel Fibroblastic Cell Line (SCI13D) Derived from the Broncho-Alveolar Lavage of a Patient with Fibrotic Hypersensitivity Pneumonitis" Biomedicines 9, no. 9: 1193. https://doi.org/10.3390/biomedicines9091193