Decellularization of Extracellular Matrix from Equine Skeletal Muscle.

Autor: Miranda CMFC; Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada; Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil. Electronic address: carlacarvalhovet@gmail.com., Therrien J; Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada., Leonel LCPC; Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil., Smith OE; Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada., Miglino MA; Department of Surgery, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, SP, Brazil., Smith LC; Centre de recherche en reproduction et fertilité, Faculté de médecine vétérinaire, Université de Montréal, Saint-Hyacinthe, Quebec, Canada.
Jazyk: angličtina
Zdroj: Journal of equine veterinary science [J Equine Vet Sci] 2020 Jul; Vol. 90, pp. 102962. Date of Electronic Publication: 2020 Mar 19.
DOI: 10.1016/j.jevs.2020.102962
Abstrakt: Equine represents an attractive animal model for musculoskeletal tissue diseases, exhibiting much similarity to the injuries that occur in humans. Cell therapy and tissue bioengineering have been widely used as a therapeutic alternative by regenerative medicine in musculoskeletal diseases. Thus, the aim of this study was to produce an acellular biomaterial of equine skeletal muscle and to evaluate its effectiveness in supporting the in vitro culture of equine induced pluripotency stem cells (iPSCs). Biceps femoris samples were frozen at -20°C for 4 days and incubated in 1% sodium dodecyl sulfate (SDS), 5 mM EDTA + 50 mM Tris and 1% Triton X-100; the effectiveness of the decellularization was evaluated by the absence of remnant nuclei (histological and 4',6-diamidino-2-phenylindole [DAPI] analysis), preservation of extracellular matrix (ECM) proteins (immunofluorescence and immunohistochemistry) and organization of ECM ultrastructure (scanning electron microscopy). Decellularized samples were recellularized with iPSCs at the concentration of 50,000 cells/cm 2 and cultured in vitro for 9 days, and the presence of the cells in the biomaterial was evaluated by histological analysis and presence of nuclei. Decellularized biomaterial showed absence of remnant nuclei and muscle fibers, as well as the preservation of ECM architecture, vascular network and proteins, laminin, fibronectin, elastin, collagen III and IV. After cellularization, iPSC nuclei were present at 9 days after incubation, indicating the decellularized biomaterial-supported iPSC survival. It is concluded that the ECM biomaterial produced from the decellularized equine skeletal muscle has potential for iPSC adhesion, representing a promising biomaterial for regenerative medicine in the therapy of musculoskeletal diseases.
(Copyright © 2020 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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