title: 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.
authors.affiliation: 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.
authors.forename: Soo-Eun
authors.initials: SE
authors.lastname: Sung
authors.affiliation: 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.
authors.forename: Meeyul
authors.initials: M
authors.lastname: Hwang
authors.affiliation: 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.
authors.forename: Ah-Young
authors.initials: AY
authors.lastname: Kim
authors.affiliation: 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.
authors.forename: Eun-Mi
authors.initials: EM
authors.lastname: Lee
authors.affiliation: 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.
authors.forename: Eun-Joo
authors.initials: EJ
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, Kyungpook National University Hospital, Daegu, Republic of Korea.
authors.forename: Su-Kyeong
authors.initials: SK
authors.lastname: Hwang
authors.affiliation: Skeletal Diseases Genome Research Center, Kyungpook National University Hospital, Daegu, Republic of Korea.
authors.forename: Shin-Yoon
authors.initials: SY
authors.lastname: Kim
authors.affiliation: Department of Ophthalmology, Kyungpook National University Hospital, Daegu, Republic of Korea.
authors.forename: Hong-Kyun
authors.initials: HK
authors.lastname: Kim
authors.affiliation: 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.
authors.forename: Kyu-Shik
authors.initials: KS
authors.lastname: Jeong
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D016189:Dystrophin / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006736:Horses; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D024510:Muscle Development / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D018008:Myogenin / Q000235:genetics / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D019210:Troponin I / Q000378:metabolism
chemical_list: D016189:Dystrophin; D017570:MyoD Protein; D018008:Myogenin; D019210:Troponin I; D018995:Myosin Heavy Chains
keywords: Adipose-derived stem cells (ADSCs); Muscle regeneration therapy; MyoD; Myogenic differentiation