MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential.
Abstract: Mesenchymal stem cells could potentially be used in the clinical treatment of muscle disorders and muscle regeneration. Adipose-derived stem cells (ADSCs) can be easily isolated from adipose tissue, as opposed to stem cells of other tissues. We believe that cell therapy using ADSCs could be applied to muscle disorders in horses and other species. We sought to improve the myogenic differentiation potential of equine ADSCs (eqADSCs) using a MyoD lentiviral vector. MyoD lentiviruses were transduced into eqADSCs and selected using puromycin. Cells were cultured in differentiation media containing 5% horse serum, and after 5 days the MyoD-transduced cells differentiated into myogenic cells (MyoD-eqADSCs). Using green fluorescent protein (GFP), MyoD-eqADSCs were purified and transplanted into the tibialis anterior muscles of mice after they were injured with the myotoxin notexin. The mice were sacrificed to examine any regeneration in the tibialis anterior muscle 4 weeks after the MyoD-eqADSCs were injected. The MyoD-eqADSCs cultured in growth media expressed murine and equine MyoD; however, they did not express late differentiation markers such as myogenin (MYOG). When cells were grown in differentiation media, the expression of MYOG was clearly observed. According to our reverse transcription polymerase chain reaction and immunocytochemistry results, MyoD-eqADSCs expressed terminal myogenic phase genes, such as those encoding dystrophin, myosin heavy chain, and troponin I. The MyoD-eqADSCs fused to each other, and the formation of myotube-like cells from myoblasts in differentiation media occurred between days 5 and 14 postplating. In mice, we observed GFP-positive myofibers, which had differentiated from the injected MyoD-eqADSCs. Our approaches improved the myogenic differentiation of eqADSCs through the forced expression of murine MyoD. Our findings suggest that limitations in the treatment of equine muscle disorders could be overcome using ADSCs.
Publication Date: 2016-02-20 PubMed ID: 26892394DOI: 10.3727/096368916X691015Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article explores the potential of using Mesenchymal stem cells, particularly Adipose-derived stem cells (ADSCs), for the treatment of muscle disorders and muscle regeneration. Their approach improves myogenic differentiation – the process of stem cells turning into muscle cells – with the aid of a MyoD lentiviral vector.
Understanding the methodology
- The researchers extracted adipose-derived stem cells (ADSCs) from equines. These stem cells have the potential to be transformed into cells of various types, in this case, the researchers focus on their differentiation into muscle cells – a process called Myogenesis.
- They used a MyoD lentiviral vector to enhance the myogenic differentiation potential of the equine ADSCs. MyoD is a protein that plays a crucial role in muscle differentiation, and lentiviral vectors are a type of virus used to deliver genes or proteins into cells. So, they used the lentiviral vector to introduce and overexpress the MyoD in ADSCs and selected the successfully transduced cells using puromycin – an antibiotic used for cell selection.
- The cells were then cultured in differentiation media for 5 days, after which the MyoD-transduced cells differentiated into myogenic cells.
Investigation
- The researchers then purified these MyoD-transduced cells using green fluorescent protein (GFP) and transplanted them into mice’s tibialis anterior muscles, previously injured with the myotoxin notexin.
- Four weeks after the transplantation, the mice were sacrificed in order to study the regeneration in the tibialis anterior muscle. GFP-positive myofibers were spotted, which had differentiated from the injected MyoD-transduced cells, indicating successful myogenesis.
- The MyoD-transduced cells also exhibited the expression of terminal myogenic phase genes, such as dystrophin, myosin heavy chain, and troponin I, confirming successful myogenic differentiation.
- These cells also exhibited the formation of myotube-like cells, indicating the progression and maturity of myogenesis.
Conclusion
- This research suggests that the limitations currently present in the treatment of equine muscle disorders, and potentially in other muscle related disorders, could be overcome with the use of adipose-derived stem cells.
- The targeted introduction and over-expression of MyoD in these cells greatly improves their myogenic differentiation, making it a promising therapeutic candidate.
Cite This Article
APA
Sung SE, Hwang M, Kim AY, Lee EM, Lee EJ, Hwang SK, Kim SY, Kim HK, Jeong KS.
(2016).
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential.
Cell Transplant, 25(11), 2017-2026.
https://doi.org/10.3727/096368916X691015 Publication
Researcher Affiliations
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
- Department of Pediatrics, Kyungpook National University Hospital, Daegu, Republic of Korea.
- Skeletal Diseases Genome Research Center, Kyungpook National University Hospital, Daegu, Republic of Korea.
- Department of Ophthalmology, Kyungpook National University Hospital, Daegu, Republic of Korea.
- Department of Pathology, College of Veterinary Medicine, Kyungpook National University, Daegu, Republic of Korea.
- Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of Korea.
MeSH Terms
- Adipose Tissue / cytology
- Animals
- Cell Differentiation / physiology
- Cells, Cultured
- Dystrophin / metabolism
- Genetic Vectors / genetics
- Genetic Vectors / metabolism
- Horses
- Lentivirus / genetics
- Mice
- Mice, Inbred BALB C
- Mice, Nude
- Muscle Development / physiology
- Muscle Fibers, Skeletal / cytology
- Muscle Fibers, Skeletal / metabolism
- MyoD Protein / genetics
- MyoD Protein / metabolism
- Myoblasts / cytology
- Myoblasts / metabolism
- Myogenin / genetics
- Myogenin / metabolism
- Myosin Heavy Chains / metabolism
- Stem Cell Transplantation
- Stem Cells / cytology
- Stem Cells / metabolism
- Troponin I / metabolism
Citations
This article has been cited 13 times.- Kankala RK, Han YH, Xia HY, Wang SB, Chen AZ. Nanoarchitectured prototypes of mesoporous silica nanoparticles for innovative biomedical applications. J Nanobiotechnology 2022 Mar 12;20(1):126.
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