Generation of Functional Myocytes from Equine Induced Pluripotent Stem Cells

Abstract

Induced pluripotent stem cells (iPSCs) have revolutionized human biomedicine through their use in disease modeling and therapy. In comparison, little progress has been made toward the application of iPSCs in veterinary species. In that regard, skeletal myocytes from iPSCs would have great potential for understanding muscle function and disease in the equine athlete. In this study, we generated skeletal myotubes by transducing equine iPSC-derived mesenchymal derivatives with an inducible lentiviral vector coding for the human sequence of the myogenic factor, MyoD. Myosin heavy chain-positive myotubes generated from two different iPSC lines were compared to myotubes from adult equine skeletal muscle progenitor cells (MPCs). iPSC myotubes had a smaller mean area than MPC myotubes (≤2-fold). In addition, quantitative polymerase chain reaction analyses showed that iPSC myotubes expressed MYH2 and MYH3 isoforms (at similar or lower levels than MPC myotubes), but they did not express the mature muscle isoform, MYH1. Compared to MPC myotubes, iPSC myotubes expressed reduced levels of the myogenic factors, MYOD1 and MYF6, but did not express MYF5. Finally, iPSC myotubes responded to KCl-induced membrane depolarization by releasing calcium and did so in a manner similar to MPC myotubes. In conclusion, this is the first study to report the generation of functional myocytes from equine iPSCs.

Induced pluripotent stem cells throughout the animal kingdom: Availability and applications

“…Furthermore, Aguiar et al[116] showed that eiPSCs could be differentiated into keratinocytes focusing on skin trauma and wound management. Other research groups have studied eventual uses for mesenchymal-like progenitors capable of chondrogenesis and adipogenesis[117] or even induction into functional osteoblasts[118] and transgenic induced myocytes[119], thus providing extra cell sources for regenerative veterinary medicine. Although some studies have already tested the potential and applicability of eiPSCs as seen above, there is still a long road ahead until eiPSCs and their derivatives are completely understood and deemed safe to use in disease models and regenerative veterinary medicine.…”

Differentiation of equine induced pluripotent stem cells into mesenchymal lineage for therapeutic use

“…Research on horses is an important aspect of veterinary medicine, particularly in the field of sports medicine, and E-iPSCs for potential injury rehabilitation have been established by our group and many other research teams [7,9,16,25,26]. In this study, E-iPSCs were differentiated into MSCs for cell transplantation to treat musculoskeletal injuries in horses (Figure 8).…”

“…In addition to their promise for use in veterinary medicine, E-iPSCs have opened up the possibility of using horses for validating stem cell-based therapies before their application in human clinical settings. Research on the differentiation of iPSCs into other cell types, including osteoblasts and myocytes, is ongoing, in efforts to advance the field of regenerative medicine for application to musculoskeletal injuries [15,16].…”

The use of induced pluripotent stem cells in domestic animals: a narrative review

“…neural progenitors [41,76,121]), to fully differentiated cell types (e.g. skeletal myocytes [122]). Domestic animal diseases are abundant and have negative health effects for consumers of agricultural animal by-products [123][124][125][126].…”

“…Equine iPSCs (eiPSCs) have been differentiated into several cell and tissue types for disease modelling including neurons [ 86 , 127 ], tendons [ 128 ], myotubes [ 122 ], and osteoblasts [ 129 ]. Functional eiPSC-derived neurons have been produced and were capable of firing action potentials in vitro via functional calcium channels [ 86 ].…”