MBP-FGF2-Immobilized Matrix Maintains Self-Renewal and Myogenic Differentiation Potential of Skeletal Muscle Stem Cells

Sah, Jay Prakash; Hao, Nguyen Thi Thu; Kim, Yunhye; Eigler, Tamar; Tzahor, Eldad; Kim, Sang‐Heon; Hwang, Yongsung; Yoon, Jong‐Hwan

doi:10.15283/ijsc18125

Cited by 4 publications

(6 citation statements)

References 20 publications

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“…Given the importance of matrix-based cues, including matrix stiffness, protein immobilization, and topographical features of cell culture substrates, with regard to cell adhesion, differentiation, as well as myoblast fusion [ 17 , 24 , 31 ], we examined whether heparin-mimicking matrix-based cues presented on the surface of GelMA-PSS hydrogels could affect the myogenic differentiation of c2c12 cells. Upon full confluence, myogenic differentiation was induced for up to 3 days using TCP, GelMA, and GelMA-PSS as cell culture substrates.…”

Section: Resultsmentioning

confidence: 99%

“…Heparin and HSPGs on the cellular surface of myoblasts have previously been shown to bind bFGF with high affinity, promoting the self-renewal of skMSCs [ 39 ]. Recently, we harnessed a bFGF-immobilized matrix as a synthetic cell culture substrate to support the cell adhesion and proliferation of skMSCs while maintaining their self-renewal capacity during in vitro expansion [ 24 ]. Similarly, we elucidated the roles of various physicochemical cues of a heparin-mimicking anionic polymer PSS, including functional groups, matrix stiffness, surface roughness, and hydrophilicity, with regard to the long-term self-renewal of hPSCs.…”

Section: Discussionmentioning

confidence: 99%

“…Molecular dynamics within focal adhesions have been shown to regulate cellular adhesion through integrins and the ECM, with these biomechanical signals being transmitted in a bidirectional manner [ 19 , 20 ]. In addition, physicochemical interactions between cells and their microenvironment consistently provoke intercellular signaling, in turn modulating gene expression and diverse signal transduction [ 21 , 22 , 23 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

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Heparin-Mimicking Polymer-Based In Vitro Platform Recapitulates In Vivo Muscle Atrophy Phenotypes

Kim

Jeong

Fendereski

et al. 2021

IJMS

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The cell–cell/cell–matrix interactions between myoblasts and their extracellular microenvironment have been shown to play a crucial role in the regulation of in vitro myogenic differentiation and in vivo skeletal muscle regeneration. In this study, by harnessing the heparin-mimicking polymer, poly(sodium-4-styrenesulfonate) (PSS), which has a negatively charged surface, we engineered an in vitro cell culture platform for the purpose of recapitulating in vivo muscle atrophy-like phenotypes. Our initial findings showed that heparin-mimicking moieties inhibited the fusion of mononucleated myoblasts into multinucleated myotubes, as indicated by the decreased gene and protein expression levels of myogenic factors, myotube fusion-related markers, and focal adhesion kinase (FAK). We further elucidated the underlying molecular mechanism via transcriptome analyses, observing that the insulin/PI3K/mTOR and Wnt signaling pathways were significantly downregulated by heparin-mimicking moieties through the inhibition of FAK/Cav3. Taken together, the easy-to-adapt heparin-mimicking polymer-based in vitro cell culture platform could be an attractive platform for potential applications in drug screening, providing clear readouts of changes in insulin/PI3K/mTOR and Wnt signaling pathways.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Heparin-Mimicking Polymer-Based In Vitro Platform Recapitulates In Vivo Muscle Atrophy Phenotypes

Kim

Jeong

Fendereski

et al. 2021

IJMS

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“…Previous studies have shown that a prerequisite step in myogenic differentiation of SkMSCs is the activation of MyoD [18,22,40,41]. Moreover, TGFβ1 was reported to regulate the ERK signaling pathway involved in in ammation, regeneration and differentiation [42,43].…”

Section: Discussionmentioning

confidence: 99%

TGFβ1 Improves the Proliferation of Decellularized and Aligned Skeletal Muscle in Extracellular Matrix Hydrogels by m6A Modification-Mediated Integrin Through Inducing ERK Phosphorylation

Zhu

Qin

Yang

et al. 2021

Preprint

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Background: The purpose of this study was to illustrate the characteristics of skeletal muscle ECM hydrogels and assess the function and underlying mechanisms of hydrogels combined with TGFβ1 for the myogenesis of skeletal muscle stem cells (SkMSCs). Methods: Six methods were used to obtain extracellular matrix (ECM) from rat skeletal muscle. Hydrogel components and cytocompatibility were assessed by Masson’s trichrome staining, scanning electron microscopy, the sulfated glycosaminoglycan (s-GAG) and bFGF contents and a cell viability assay. Satellite cells were sorted, identified and seeded into skeletal muscle ECM hydrogels with TGFβ1. To determine the function of decellularized aligned skeletal muscle ECM hydrogels with TGFβ1 for the proliferation and differentiation of SkMSCs, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 5-ethynyl-2′-deoxyuridine (EdU) analysis and immunofluorescence staining were conducted. RNA stability assay and Gene set enrichment analysis (GSEA) were used to further investigated the mechanism underlying the function. Results: Our study established a highly efficient method for the decellularization of skeletal muscle and fabricated an ECM hydrogel without cell components that preserves the rich ECM composition. This hydrogel formed a three-dimensional microstructure and showed good biocompatibility. We found that the decellularized aligned ECM scaffold with TGFβ1 promoted SkMSC proliferation and suppressed SkMSC differentiation through extracellular signal-regulated kinase (ERK) signaling. Our study suggested that TGFβ1 increases the m6A methylation of the integrin mRNA 3’UTR, thereby inducing the phosphorylation of ERK and promoting SkMSCs proliferation and differentiation.Conclusions: Our study explored a highly efficient method for the decellularization of skeletal muscle and fabricated an ECM hydrogel with a three-dimensional microstructure and good biocompatibility. Furthermore, we demonstrated that decellularized aligned ECM scaffolds with TGFβ1 promoted SkMSC proliferation and differentiation through enhancing the m6A methylation of the integrin mRNA 3’UTR, which may serve as a potential therapeutic strategy for the acute and chronic muscle injuries

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“…Estos resultados son contradictorios ya que otros autores como Sah et al (2019) han utilizado el FGF-2 en la elaboración de matrices sintéticas que permiten conservar el fenotipo inicial de las CM de músculo esquelético; las células cultivadas en dicha matriz fueron aisladas de los músculos de las extremidades posteriores de ratones ICR (Institute of Cancer Research) de 6 a 8 semanas de edad y sembradas en medio de crecimiento (Ham-F10 con suero de caballo al 20 %, 1 % penicilina-estreptomicina, 5 ng/ml de FGF2 humano, en una incubadora con 5 % de CO 2 a 37 • C), registrando un estado indiferenciado de las células cultivadas en dicha matriz y demostrando una fuerte capacidad de proliferación. Este estudio fue corroborado por los análisis de Endo et al (2019).…”

Section: Discussionunclassified

Influencia del factor de crecimiento fibroblástico 2 en células madre in vitro

Vargas¹,

Arzuza²,

Turner³

2020

Actual. Biol.

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El interés y la necesidad de estudiar las células madre en terapias regenerativas han aumentado en los últimos años debido a su capacidad de proliferación y diferenciación hacia múltiples linajes. Desafortunadamente, el número de células demandado para un trasplante satisfactorio es mayor al que se logra extraer directamente del paciente, por lo que se deben realizar cultivos in vitro de células madre. Sin embargo, con el tiempo las células de estos cultivos se vuelven senescentes, disminuyendo así su número de divisiones celulares y su capacidad proliferativa. Para solucionar esta problemática se han propuesto factores de crecimiento como agentes potenciadores de la proliferación celular. Entre estos se encuentra el factor de crecimiento fibroblástico 2, que al agregarse al medio podría promover la proliferación y aumentar el tiempo de vida de las células en cultivo. En la siguiente revisión se recopila información sobre la biología de este factor de crecimiento y el tipo de señalización que utiliza, al igual que sus aplicaciones en terapia regenerativa y su efecto en la proliferación celular y senescencia.

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MBP-FGF2-Immobilized Matrix Maintains Self-Renewal and Myogenic Differentiation Potential of Skeletal Muscle Stem Cells

Cited by 4 publications

References 20 publications

Heparin-Mimicking Polymer-Based In Vitro Platform Recapitulates In Vivo Muscle Atrophy Phenotypes

Heparin-Mimicking Polymer-Based In Vitro Platform Recapitulates In Vivo Muscle Atrophy Phenotypes

TGFβ1 Improves the Proliferation of Decellularized and Aligned Skeletal Muscle in Extracellular Matrix Hydrogels by m6A Modification-Mediated Integrin Through Inducing ERK Phosphorylation

Influencia del factor de crecimiento fibroblástico 2 en células madre in vitro

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