Myogenic Differentiation of Human Myoblasts and Mesenchymal Stromal Stem Cells under GDF11 on Poly-ɛ-Caprolactone-Collagen I-Polyethylene-Nanofibers

Abstract Primary myoblasts (Mb) and adipose derived mesenchymal stromal cells (ADSC) can be co-cultured and myogenically differentiated in the process of skeletal muscle tissue engineering. Electrospun composite nanofiber scaffolds represent suitable matrices for tissue engineering of skeletal muscle, combining biocompatibility and stability. Although growth differentiation factor 11 (GDF11) has been proposed as a rejuvenating circulating factor, restoring skeletal muscle function in aging mice, some studies have also described a harming effect of GDF11.Therefore the aim of the study was to analyze the effect of GDF11 on co-cultures of Mb and ADSC on poly-ε-caprolacton (PCL)-collagen I-polyethylene oxide (PEO)-nanofibers.Human Mb were co-cultured with ADSC two-dimensionally (2D) as monolayers or three-dimensionally (3D) on aligned PCL-collagen I-PEO-nanofibers. Differentiation media were either serum-free with or without GDF11, or serum containing as in a conventional differentiation medium. Cell viability was higher after conventional myogenic differentiation compared to serum-free and serum-free + GDF11 differentiation as was creatine kinase activity. Immunofluorescence staining showed myosin heavy chain expression in all groups after 28 days of differentiation. Gene expression of myosin heavy chain (MYH2) increased after serum-free + GDF11 stimulation compared to serum-free stimulation alone. The results of this study show that PCL-collagen I-PEO-nanofibers represent a suitable matrix for 3D myogenic differentiation of Mb and ADSC. In this context, GDF11 seems to promote myogenic differentiation of Mb and ADSC co-cultures compared to serum-free differentiation without any evidence of a harming effect.

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ProNGF/p75NTR Axis Drives Fiber Type Specification by Inducing the Fast-Glycolytic Phenotype in Mouse Skeletal Muscle Cells

Cells ◽

10.3390/cells9102232 ◽

2020 ◽

Vol 9 (10) ◽

pp. 2232

Author(s):

Valentina Pallottini ◽

Mayra Colardo ◽

Claudia Tonini ◽

Noemi Martella ◽

Georgios Strimpakos ◽

...

Keyword(s):

Skeletal Muscle ◽

Myosin Heavy Chain ◽

Heavy Chain ◽

Myogenic Differentiation ◽

Fiber Type ◽

Pcr Analysis ◽

Mdx Mouse ◽

Mdx Mice ◽

Muscle Physiology

Despite its undisputable role in the homeostatic regulation of the nervous system, the nerve growth factor (NGF) also governs the relevant cellular processes in other tissues and organs. In this study, we aimed at assessing the expression and the putative involvement of NGF signaling in skeletal muscle physiology. To reach this objective, we employed satellite cell-derived myoblasts as an in vitro culture model. In vivo experiments were performed on Tibialis anterior from wild-type mice and an mdx mouse model of Duchenne muscular dystrophy. Targets of interest were mainly assessed by means of morphological, Western blot and qRT-PCR analysis. The results show that proNGF is involved in myogenic differentiation. Importantly, the proNGF/p75NTR pathway orchestrates a slow-to-fast fiber type transition by counteracting the expression of slow myosin heavy chain and that of oxidative markers. Concurrently, proNGF/p75NTR activation facilitates the induction of fast myosin heavy chain and of fast/glycolytic markers. Furthermore, we also provided evidence that the oxidative metabolism is impaired in mdx mice, and that these alterations are paralleled by a prominent buildup of proNGF and p75NTR. These findings underline that the proNGF/p75NTR pathway may play a crucial role in fiber type determination and suggest its prospective modulation as an innovative therapeutic approach to counteract muscle disorders.

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Myogenic differentiation of primary myoblasts and mesenchymal stromal cells under serum-free conditions on PCL-collagen I-nanoscaffolds

BMC Biotechnology ◽

10.1186/s12896-018-0482-6 ◽

2018 ◽

Vol 18 (1) ◽

Cited By ~ 4

Author(s):

Aijia Cai ◽

Moritz Hardt ◽

Paul Schneider ◽

Rafael Schmid ◽

Claudia Lange ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Myogenic Differentiation ◽

Collagen I ◽

Serum Free ◽

Primary Myoblasts ◽

Serum Free Conditions

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A Calcineurin-NFATc3-Dependent Pathway Regulates Skeletal Muscle Differentiation and Slow Myosin Heavy-Chain Expression

Molecular and Cellular Biology ◽

10.1128/mcb.20.17.6600-6611.2000 ◽

2000 ◽

Vol 20 (17) ◽

pp. 6600-6611 ◽

Cited By ~ 211

Author(s):

Ulrike Delling ◽

Jolana Tureckova ◽

Hae W. Lim ◽

Leon J. De Windt ◽

Peter Rotwein ◽

...

Keyword(s):

Skeletal Muscle ◽

Signal Transduction ◽

Gene Transfer ◽

Myosin Heavy Chain ◽

Heavy Chain ◽

Myogenic Differentiation ◽

Fiber Type ◽

Slow Myosin ◽

Slow Myosin Heavy Chain ◽

Myocyte Differentiation

ABSTRACT The differentiation and maturation of skeletal muscle cells into functional fibers is coordinated largely by inductive signals which act through discrete intracellular signal transduction pathways. Recently, the calcium-activated phosphatase calcineurin (PP2B) and the family of transcription factors known as NFAT have been implicated in the regulation of myocyte hypertrophy and fiber type specificity. Here we present an analysis of the intracellular mechanisms which underlie myocyte differentiation and fiber type specificity due to an insulinlike growth factor 1 (IGF-1)–calcineurin–NFAT signal transduction pathway. We demonstrate that calcineurin enzymatic activity is transiently increased during the initiation of myogenic differentiation in cultured C2C12 cells and that this increase is associated with NFATc3 nuclear translocation. Adenovirus-mediated gene transfer of an activated calcineurin protein (AdCnA) potentiates C2C12 and Sol8 myocyte differentiation, while adenovirus-mediated gene transfer of noncompetitive calcineurin-inhibitory peptides (cain or ΔAKAP79) attenuates differentiation. AdCnA infection was also sufficient to rescue myocyte differentiation in an IGF-depleted myoblast cell line. Using 10T1/2 cells, we demonstrate that MyoD-directed myogenesis is dramatically enhanced by either calcineurin or NFATc3 cotransfection, while a calcineurin inhibitory peptide (cain) blocks differentiation. Enhanced myogenic differentiation directed by calcineurin, but not NFATc3, preferentially specifies slow myosin heavy-chain expression, while enhanced differentiation through mitogen-activated protein kinase kinase 6 (MKK6) promotes fast myosin heavy-chain expression. These data indicate that a signaling pathway involving IGF-calcineurin-NFATc3 enhances myogenic differentiation whereas calcineurin acts through other factors to promote the slow fiber type program.

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Skeletal muscle tissue engineering: A maturation model promoting long-term survival of myotubes, structural development of the excitation–contraction coupling apparatus and neonatal myosin heavy chain expression

Biomaterials ◽

10.1016/j.biomaterials.2009.05.081 ◽

2009 ◽

Vol 30 (29) ◽

pp. 5392-5402 ◽

Cited By ~ 31

Author(s):

Mainak Das ◽

John W. Rumsey ◽

Neelima Bhargava ◽

Maria Stancescu ◽

James J. Hickman

Keyword(s):

Skeletal Muscle ◽

Tissue Engineering ◽

Myosin Heavy Chain ◽

Muscle Tissue ◽

Heavy Chain ◽

Structural Development ◽

Term Survival ◽

Excitation Contraction Coupling ◽

Contraction Coupling

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17β-Estradiol regulates the first steps of skeletal muscle cell differentiation via ER-α-mediated signals

AJP Cell Physiology ◽

10.1152/ajpcell.00188.2009 ◽

2009 ◽

Vol 297 (5) ◽

pp. C1249-C1262 ◽

Cited By ~ 45

Author(s):

Paola Galluzzo ◽

Chiara Rastelli ◽

Pamela Bulzomi ◽

Filippo Acconcia ◽

Valentina Pallottini ◽

...

Keyword(s):

Skeletal Muscle ◽

Myosin Heavy Chain ◽

Heavy Chain ◽

Glucose Transporter ◽

Reproductive Tract ◽

Myogenic Differentiation ◽

Trophic Factors ◽

Differentiation Markers ◽

17Β Estradiol ◽

Myoblast Cells

17β-Estradiol (E2) mediates a wide variety of complex biological processes determining the growth and development of reproductive tract as well as nonreproductive tissues of male and female individuals. While E2 effects on the reproductive system, bone, and cardiovascular system are quite well established, less is known about how it affects the physiology of other tissues. Skeletal muscle is a tissue that is expected to be E2 responsive since both isoforms of estrogen receptor (ER-α and ER-β) are expressed. Significant sex-related differences have been described in skeletal muscle, although the role played by E2 and the mechanisms underlying it remain to be determined. Here, we demonstrate that E2 increases the glucose transporter type 4 translocation at membranes as well as the expression of well-known differentiation markers of myogenesis (i.e., myogenin and myosin heavy chain) in rat myoblast cells (L6). These E2-induced effects require rapid extranuclear signals and the presence of ER-α, whereas no contribution of IGF-I receptor has been observed. In particular, ER-α-dependent Akt activation participates in regulating the first step of myogenic differentiation. Moreover, both receptors mediate the E2-induced activation of p38, which, in turn, affects the expression of myogenin and myosin heavy chain. All together, these data indicate that E2 should be included in the list of skeletal muscle trophic factors.

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