Mesenchymal stem cells and myoblast differentiation under HGF and IGF-1 stimulation for 3D skeletal muscle tissue engineering

Abstract

BackgroundVolumetric muscle loss caused by trauma or after tumour surgery exceeds the natural regeneration capacity of skeletal muscle. Hence, the future goal of tissue engineering (TE) is the replacement and repair of lost muscle tissue by newly generating skeletal muscle combining different cell sources, such as myoblasts and mesenchymal stem cells (MSCs), within a three-dimensional matrix. Latest research showed that seeding skeletal muscle cells on aligned constructs enhance the formation of myotubes as well as cell alignment and may provide a further step towards the clinical application of engineered skeletal muscle.In this study the myogenic differentiation potential of MSCs upon co-cultivation with myoblasts and under stimulation with hepatocyte growth factor (HGF) and insulin-like growth factor-1 (IGF-1) was evaluated. We further analysed the behaviour of MSC-myoblast co-cultures in different 3D matrices.ResultsPrimary rat myoblasts and rat MSCs were mono- and co-cultivated for 2, 7 or 14 days. The effect of different concentrations of HGF and IGF-1 alone, as well as in combination, on myogenic differentiation was analysed using microscopy, multicolour flow cytometry and real-time PCR. Furthermore, the influence of different three-dimensional culture models, such as fibrin, fibrin-collagen-I gels and parallel aligned electrospun poly-ε-caprolacton collagen-I nanofibers, on myogenic differentiation was analysed. MSCs could be successfully differentiated into the myogenic lineage both in mono- and in co-cultures independent of HGF and IGF-1 stimulation by expressing desmin, myocyte enhancer factor 2, myosin heavy chain 2 and alpha-sarcomeric actinin. An increased expression of different myogenic key markers could be observed under HGF and IGF-1 stimulation. Even though, stimulation with HGF/IGF-1 does not seem essential for sufficient myogenic differentiation. Three-dimensional cultivation in fibrin-collagen-I gels induced higher levels of myogenic differentiation compared with two-dimensional experiments. Cultivation on poly-ε-caprolacton-collagen-I nanofibers induced parallel alignment of cells and positive expression of desmin.ConclusionsIn this study, we were able to myogenically differentiate MSC upon mono- and co-cultivation with myoblasts. The addition of HGF/IGF-1 might not be essential for achieving successful myogenic differentiation. Furthermore, with the development of a biocompatible nanofiber scaffold we established the basis for further experiments aiming at the generation of functional muscle tissue.Electronic supplementary materialThe online version of this article (doi:10.1186/s12860-017-0131-2) contains supplementary material, which is available to authorized users.

Improved therapeutics of modified mesenchymal stem cells: an update

“…Hepatocyte growth factor (HGF) is another significant cytokine that participate in angiogenesis, anti-inflammation and anti-apoptosis. MSCs overexpressing HGF are reported to have high therapeutic influence in cardiovascular diseases [54], liver injuries [55], lungs conditions [56], stabilizing endothelial barrier function [57,58], skeletal muscle tissue regeneration [59], and even regeneration of damaged neurons in a Parkinson's disease model [60]. Jang and colleagues investigate the protective role of HGF gene-transfected MSCs in acetaminophen induced hepatocytes injury.…”

Five Decades Later, Are Mesenchymal Stem Cells Still Relevant?

“…Such matrices can be worked into desired shapes using micromolding, microfluidics, electrostatic droplet extrusion, or bioprinting (Kim et al, 2019). MSC based scaffolds systems have been used for bone and cartilage regeneration (Kim et al, 2019), as well as for the reproduction of blood vessels (Pinnock et al, 2016), cardiac tissue (Rashedi et al, 2017;Ichihara et al, 2018;Joshi et al, 2018), and skeletal muscle (Witt et al, 2017). Optimal tissue replacement efficiency relies on the physical characteristics of the scaffolds (Alakpa et al, 2017;Jeon et al, 2017;Mouser et al, 2018), as each mechanical property can modify the fate of the transplanted cells.…”

Biofabrikation – neue Ansätze für den artifiziellen Gewebeersatz

“…Diese werden zur Ver vielfältigung meist im Labor kultiviert, um sie dann in den Emp fängerorganismus zu implantieren [6][7][8][9][10][11][12]. In vitro unter Laborbe dingungen und im Kleintiermodell [13] hat diese Methode teils erstaunliche Ergebnisse gezeigt [14][15][16]. U.a.…”