Abstract
During development, cardiac and skeletal muscle share major transcription factors and sarcomere proteins which were generally regarded as specific to either cardiac or skeletal muscle but not both in terminally differentiated adult cardiac or skeletal muscle. Here, we investigated whether artificial muscle constructed from human skeletal muscle derived stem cells (MDSCs) recapitulates developmental similarities between cardiac and skeletal muscle. We constructed 3-dimensional collagen-based engineered muscle tissue (EMT) using MDSCs (MDSC-EMT) and compared the biochemical and contractile properties with EMT using induced pluripotent stem (iPS) cell-derived cardiac cells (iPS-EMT). Both MDSC-EMT and iPS-EMT expressed cardiac specific troponins, fast skeletal muscle myosin heavy chain, and connexin-43 mimicking developing cardiac or skeletal muscle. At the transcriptional level, MDSC-EMT and iPS-EMT upregulated both cardiac and skeletal muscle-specific genes and expressed Nkx2.5 and Myo-D proteins. MDSC-EMT displayed intracellular calcium ion transients and responses to isoproterenol. Contractile force measurements of MDSC-EMT demonstrated functional properties of immature cardiac and skeletal muscle in both tissues. Results suggest that the EMT from MDSCs mimics developing cardiac and skeletal muscle and can serve as a useful in vitro functioning striated muscle model for investigation of stem cell differentiation and therapeutic options of MDSCs for cardiac repair.
Highlights
The adult heart is largely a nonregenerative organ
Our results indicate that muscle derived stem cells (MDSCs)-engineered muscle tissue (EMT) and induced pluripotent stem (iPS)-EMT share a number of biochemical similarities, but iPS-EMT has a better degree of electrical coupling and adrenergic responsiveness
Human Y1 iPS cells were previously established in Lei Yang’s lab from healthy fibroblasts [10] and used as the control. iPS cells were maintained on mouse embryonic fibroblast (MEF) feeder layers with regular human embryonic stem cell medium containing 10 ng/mL FGF-2 [9]. iPS cells were differentiated into cardiomyocytes using our previously established protocol [9, 10]
Summary
The adult heart is largely a nonregenerative organ. Cardiomyocytes (CMs), the contractile cells of the heart, have a modest rate of turnover, ranging from 1% in youth to less than 0.5% in old age [1], this level is not enough to compensate for the large number of cardiomyocytes which are lost as a result of heart injury. MDSCs can be rapidly expanded in vitro to obtain clinically relevant numbers of cells, which can be transplanted as an autologous graft. They are advantageous because they are resistant to hypoxia, attenuate fibrosis, and readily differentiate into contractile cells [4]. We previously showed that rodent MDSCs differentiate into CM-like cells with cardiaclike electrophysiological, biochemical, and contractile properties using cell aggregate formation and 3-dimensional (3D)
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