Impaired primary mouse myotube formation on crosslinked type I collagen films is enhanced by laminin and entactin

Grefte, Sander; Adjobo‐Hermans, Merel J. W.; Versteeg, E.M.M.; Koopman, Werner J.H.; Daamen, Willeke F.

doi:10.1016/j.actbio.2015.11.009

Cited by 16 publications

(11 citation statements)

References 79 publications

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“…In agreement with this finding, studies have shown that both mouse C2C12 and human skeletal myotube formation is enhanced on laminin-μprinted surfaces compared to surfaces μprinted with fibronectin, Type I collagen, or Type IV collagen 13 . Similarly, myoblast adhesion to Collagen I films was shown to be enhanced by coating the films with laminin 31 . These studies together suggest that laminin is a strong cue for myotube formation and adhesion, which was lacking from our constructs.…”

Section: Discussionmentioning

confidence: 93%

Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

Bettadapur

Suh

Geisse

et al. 2016

Sci Rep

111

145

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In vitro models of skeletal muscle are critically needed to elucidate disease mechanisms, identify therapeutic targets, and test drugs pre-clinically. However, culturing skeletal muscle has been challenging due to myotube delamination from synthetic culture substrates approximately one week after initiating differentiation from myoblasts. In this study, we successfully maintained aligned skeletal myotubes differentiated from C2C12 mouse skeletal myoblasts for three weeks by utilizing micromolded (μmolded) gelatin hydrogels as culture substrates, which we thoroughly characterized using atomic force microscopy (AFM). Compared to polydimethylsiloxane (PDMS) microcontact printed (μprinted) with fibronectin (FN), cell adhesion on gelatin hydrogel constructs was significantly higher one week and three weeks after initiating differentiation. Delamination from FN-μprinted PDMS precluded robust detection of myotubes. Compared to a softer blend of PDMS μprinted with FN, myogenic index, myotube width, and myotube length on μmolded gelatin hydrogels was similar one week after initiating differentiation. However, three weeks after initiating differentiation, these parameters were significantly higher on μmolded gelatin hydrogels compared to FN-μprinted soft PDMS constructs. Similar results were observed on isotropic versions of each substrate, suggesting that these findings are independent of substrate patterning. Our platform enables novel studies into skeletal muscle development and disease and chronic drug testing in vitro.

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Section: Discussionmentioning

confidence: 93%

Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

Bettadapur

Suh

Geisse

et al. 2016

Sci Rep

111

145

View full text Add to dashboard Cite

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“…The laminin-nidogen complex contains the laminins LN111 and LN211 also present in the basal lamina of the skeletal muscle tissue [13]. The concentration of 100 μg/mL we used previously showed improved myotube formation on collagen coatings in vitro [14]. A similar concentration of laminin showed improved myofiber formation in vivo in rats [25].…”

Section: Fibrin Constructsmentioning

confidence: 89%

“…These components are produced by local fibroblasts in the healthy tissue, but also by SC-derived myoblasts during muscle repair [9]. In vitro, laminin-nidogen promotes myofiber formation on collagen coatings, indicating it may also be suitable to enhance muscle constructs [14].…”

Section: Introductionmentioning

confidence: 99%

Fibrin with Laminin-Nidogen Reduces Fibrosis and Improves Soft Palate Regeneration Following Palatal Injury

Salazar

Rheden

Hulzen³

et al. 2021

Biomolecules

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This study aimed to analyze the effects of fibrin constructs enhanced with laminin-nidogen, implanted in the wounded rat soft palate. Fibrin constructs with and without laminin-nidogen were implanted in 1 mm excisional wounds in the soft palate of 9-week-old rats and compared with the wounded soft palate without implantation. Collagen deposition and myofiber formation were analyzed at days 3, 7, 28 and 56 after wounding by histochemistry. In addition, immune staining was performed for a-smooth muscle actin (a-SMA), myosin heavy chain (MyHC) and paired homeobox protein 7 (Pax7). At day 56, collagen areas were smaller in both implant groups (31.25 ± 7.73% fibrin only and 21.11 ± 6.06% fibrin with laminin-nidogen)) compared to the empty wounds (38.25 ± 8.89%, p < 0.05). Moreover, the collagen area in the fibrin with laminin-nidogen group was smaller than in the fibrin only group (p ˂ 0.05). The areas of myofiber formation in the fibrin only group (31.77 ± 10.81%) and fibrin with laminin-nidogen group (43.13 ± 10.39%) were larger than in the empty wounds (28.10 ± 11.68%, p ˂ 0.05). Fibrin-based constructs with laminin-nidogen reduce fibrosis and improve muscle regeneration in the wounded soft palate. This is a promising strategy to enhance cleft soft palate repair and other severe muscle injuries.

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“…All the reads that did not aligned during the first step were realigned with Bowtie [39] using a local alignment strategy. The counts related to each gene were computed using Htseq-count [40]. The row count matrix was normalized to take into account both the GC content biases and the different depth of coverage.…”

Section: Rna Extraction Sequencing and Analysismentioning

confidence: 99%

“…Last but not least, besides designing our microchannels with defined biomechanics and topology, we also wanted to equip them with a proper pro-adhesive and differentiating stimulus. To this aim, their inner wall was coated with laminin ( Fig 2E), as laminins are part of the basal lamina of skeletal muscle fibres and have been shown to promote myoblast adhesion and myotube formation both in vitro and in vivo [40][41][42].…”

Section: D Micro-channel Hydrogel Designmentioning

confidence: 99%

Engineering a 3D in vitro model of human skeletal muscle at the single fiber scale

et al. 2020

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The reproduction of reliable in vitro models of human skeletal muscle is made harder by the intrinsic 3D structural complexity of this tissue. Here we coupled engineered hydrogel with 3D structural cues and specific mechanical properties to derive human 3D muscle constructs ("myobundles") at the scale of single fibers, by using primary myoblasts or myoblasts derived from embryonic stem cells. To this aim, cell culture was performed in confined, laminin-coated micrometric channels obtained inside a 3D hydrogel characterized by the optimal stiffness for skeletal muscle myogenesis. Primary myoblasts cultured in our 3D culture system were able to undergo myotube differentiation and maturation, as demonstrated by the proper expression and localization of key components of the sarcomere and sarcolemma. Such approach allowed the generation of human myobundles of~10 mm in length and 120 μm in diameter, showing spontaneous contraction 7 days after cell seeding. Transcriptome analyses showed higher similarity between 3D myobundles and skeletal signature, compared to that found between 2D myotubes and skeletal muscle, mainly resulting from expression in 3D myobundles of categories of genes involved in skeletal muscle maturation, including extracellular matrix organization. Moreover, imaging analyses confirmed that structured 3D culture system was conducive to differentiation/maturation also when using myoblasts derived from embryonic stem cells. In conclusion, our structured 3D model is a promising tool for modelling human skeletal muscle in healthy and diseases conditions.

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Impaired primary mouse myotube formation on crosslinked type I collagen films is enhanced by laminin and entactin

Cited by 16 publications

References 79 publications

Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

Fibrin with Laminin-Nidogen Reduces Fibrosis and Improves Soft Palate Regeneration Following Palatal Injury

Engineering a 3D in vitro model of human skeletal muscle at the single fiber scale

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