Chapter title |
Test of Antifibrotic Drugs in a Cellular Model of Fibrosis Based on Muscle-Derived Fibroblasts from Duchenne Muscular Dystrophy Patients
|
---|---|
Chapter number | 15 |
Book title |
Duchenne Muscular Dystrophy
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7374-3_15 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7373-6, 978-1-4939-7374-3
|
Authors |
Simona Zanotti, Marina Mora |
Abstract |
An in vitro model of muscle fibrosis, based on the use of primary human fibroblasts isolated from muscle biopsies of patients affected by Duchenne muscular dystrophies (DMD) and cultivated in monolayer and 3D conditions, is used to test the potential antifibrotic activity of pirfenidone (PFD). This in vitro model may be usefully also to evaluate the toxicity and efficacy of other candidate molecules for the treatment of fibrosis. The drug toxicity is evaluated using a colorimetric assay based on the conversion of tetrazolium salt (MTT) to insoluble formazan, while the effect of the drug on cell proliferation is measured with the bromodeoxyuridine incorporation assay. The efficacy of the drug is evaluated in fibroblast monolayers by quantitating synthesis and deposition of intracellular collagen with a spectrophotometric picrosirius red-based assay, and by quantitating cell migration using a "scratch" assay. The efficacy of PFD as antifibrotic drug is also evaluated in a 3D fibroblast model by measuring diameters and number of nodules. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 10 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 3 | 30% |
Student > Doctoral Student | 1 | 10% |
Other | 1 | 10% |
Student > Ph. D. Student | 1 | 10% |
Student > Bachelor | 1 | 10% |
Other | 0 | 0% |
Unknown | 3 | 30% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 20% |
Pharmacology, Toxicology and Pharmaceutical Science | 1 | 10% |
Nursing and Health Professions | 1 | 10% |
Neuroscience | 1 | 10% |
Unknown | 5 | 50% |