MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential
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...
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doaj-160cad0a34b147efb8954db42ff622812020-11-25T03:17:35ZengSAGE PublishingCell Transplantation0963-68971555-38922016-11-012510.3727/096368916X691015MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation PotentialSoo-Eun Sung0Meeyul Hwang1Ah-Young Kim2Eun-Mi Lee3Eun-Joo Lee4Su-Kyeong Hwang5Shin-Yoon Kim6Hong-Kyun Kim M.D., Ph.D.7Kyu-Shik Jeong D.V.M., Ph.D.8Stem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaStem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaStem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaStem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaStem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaDepartment of Pediatrics, Kyungpook National University Hospital, Daegu, Republic of KoreaSkeletal Diseases Genome Research Center, Kyungpook National University Hospital, Daegu, Republic of KoreaDepartment of Ophthalmology, Kyungpook National University Hospital, Daegu, Republic of KoreaStem Cell Therapeutic Research Institute, Kyungpook National University, Daegu, Republic of KoreaMesenchymal 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.https://doi.org/10.3727/096368916X691015 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Soo-Eun Sung Meeyul Hwang Ah-Young Kim Eun-Mi Lee Eun-Joo Lee Su-Kyeong Hwang Shin-Yoon Kim Hong-Kyun Kim M.D., Ph.D. Kyu-Shik Jeong D.V.M., Ph.D. |
spellingShingle |
Soo-Eun Sung Meeyul Hwang Ah-Young Kim Eun-Mi Lee Eun-Joo Lee Su-Kyeong Hwang Shin-Yoon Kim Hong-Kyun Kim M.D., Ph.D. Kyu-Shik Jeong D.V.M., Ph.D. MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential Cell Transplantation |
author_facet |
Soo-Eun Sung Meeyul Hwang Ah-Young Kim Eun-Mi Lee Eun-Joo Lee Su-Kyeong Hwang Shin-Yoon Kim Hong-Kyun Kim M.D., Ph.D. Kyu-Shik Jeong D.V.M., Ph.D. |
author_sort |
Soo-Eun Sung |
title |
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential |
title_short |
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential |
title_full |
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential |
title_fullStr |
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential |
title_full_unstemmed |
MyoD Overexpressed Equine Adipose-Derived Stem Cells Enhanced Myogenic Differentiation Potential |
title_sort |
myod overexpressed equine adipose-derived stem cells enhanced myogenic differentiation potential |
publisher |
SAGE Publishing |
series |
Cell Transplantation |
issn |
0963-6897 1555-3892 |
publishDate |
2016-11-01 |
description |
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. |
url |
https://doi.org/10.3727/096368916X691015 |
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