Wharton’s Jelly-Derived Mesenchymal Stem Cells Reduce Fibrosis in a Mouse Model of Duchenne Muscular Dystrophy by Upregulating microRNA 499
Abstract
:1. Introduction
2. Materials and Methods
2.1. Ethical Statement
2.2. Animals
2.3. WJ-MSC and MMP-1 Administration
2.4. Grip Strength Measurements
2.5. CK Assays
2.6. IHC, Sirius Red Staining, and H&E Staining
2.7. PCR
2.8. miRNA Sequencing and Data Analysis
2.9. miRNA Isolation and qPCR
2.10. RNA Isolation and qPCR
2.11. Western Blot Analysis
2.12. Cell Culture
2.13. miRNA Mimic Transfection
2.14. Immunocytochemistry
2.15. Statistical Analyses
3. Results
3.1. Therapeutic Effects of WJ-MSCs in Mdx Mice
3.2. MicroRNA (miRNA) Sequencing of Mdx Mouse Skeletal Muscle Identifies Downregulation of miR-499-5p
3.3. miR-499-5p Directly Targets Transforming Growth Factor β Receptor (TGFβR)1 and TGFβR3
3.4. Anti-Fibrotic Effect of WJ-MSCs in the Skeletal Muscle of Mdx Mice
3.5. Anti-Fibrotic Effect of WJ-MSCs in the Diaphragm of Mdx Mice
3.6. Anti-Fibrotic Effect of miR-499-5p in H2O2-Treated Myotubes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Park, S.E.; Jeong, J.B.; Oh, S.J.; Kim, S.J.; Kim, H.; Choi, A.; Choi, S.-j.; Oh, S.-y.; Ryu, G.H.; Lee, J.; et al. Wharton’s Jelly-Derived Mesenchymal Stem Cells Reduce Fibrosis in a Mouse Model of Duchenne Muscular Dystrophy by Upregulating microRNA 499. Biomedicines 2021, 9, 1089. https://doi.org/10.3390/biomedicines9091089
Park SE, Jeong JB, Oh SJ, Kim SJ, Kim H, Choi A, Choi S-j, Oh S-y, Ryu GH, Lee J, et al. Wharton’s Jelly-Derived Mesenchymal Stem Cells Reduce Fibrosis in a Mouse Model of Duchenne Muscular Dystrophy by Upregulating microRNA 499. Biomedicines. 2021; 9(9):1089. https://doi.org/10.3390/biomedicines9091089
Chicago/Turabian StylePark, Sang Eon, Jang Bin Jeong, Shin Ji Oh, Sun Jeong Kim, Hyeongseop Kim, Alee Choi, Suk-joo Choi, Soo-young Oh, Gyu Ha Ryu, Jeehun Lee, and et al. 2021. "Wharton’s Jelly-Derived Mesenchymal Stem Cells Reduce Fibrosis in a Mouse Model of Duchenne Muscular Dystrophy by Upregulating microRNA 499" Biomedicines 9, no. 9: 1089. https://doi.org/10.3390/biomedicines9091089