Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

Schaft, Daisy W. J. van der; Spreeuwel, Ariane C. C. van; Boonen, Kristel J. M.; Langelaan, M.L.P.; Bouten, C.V.C.; Baaijens, F.P.T.

doi:10.3791/4267-v

Cited by 4 publications

(3 citation statements)

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“…VBAM and MDCK experiments were imaged before and after clearing. Tissue clearing was performed similar to previously published protocols (Sanchez and Morgan, 2021). Briefly, samples were dehydrated in excess methanol on a rocker at 4°C 3× for 20 min.…”

Section: Image Acquisition Tissue Clearing and Volumetric Quantificationmentioning

confidence: 99%

In vitroformation and extended culture of highly metabolically active and contractile tissues

Bagdasarian

Tonmoy

Park

et al. 2023

Preprint

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3D cell culture models have gained popularity in recent years as an alternative to animal and 2D cell culture models for pharmaceutical testing and disease modeling. Polydimethylsiloxane (PDMS) is a cost-effective and accessible molding material for 3D cultures; however, routine PDMS molding may not be appropriate for extended culture of contractile and metabolically active tissues. Failures can include loss of culture adhesion to the PDMS mold and limited culture surfaces for nutrient and waste diffusion. In this study, we evaluated PDMS molding materials and surface treatments for highly contractile and metabolically active 3D cell cultures. PDMS functionalized with polydopamine allowed for extended culture duration (14.8 ± 3.97 days) when compared to polyethylamine/glutaraldehyde functionalization (6.94 ± 2.74 days); Additionally, porous PDMS extended culture duration (16.7 ± 3.51 days) compared to smooth PDMS (6.33 ± 2.05 days) after treatment with TGF-β to increase culture contraction. Porous PDMS additionally allowed for large (13 mm tall × 8 mm diameter) constructs to be fed by diffusion through the mold, resulting in increased cell density (0.0210 ± 0.0049 mean nuclear fraction) compared to controls (0.0045 ± 0.0016 mean nuclear fraction). As a practical demonstration of the flexibility of porous PDMS, we engineered a vascular bioartificial muscle model (VBAM) and demonstrated extended culture of VBAMs anchored with porous PDMS posts. Using this model, we assessed the effect of feeding frequency on VBAM cellularity. Feeding 3×/week significantly increased nuclear fraction at multiple tissue depths relative to 2×/day. VBAM maturation was similarly improved in 3×/week feeding as measured by nuclear alignment (23.49° ± 3.644) and nuclear aspect ratio (2.274 ± 0.0643) relative to 2x/day (35.93° ± 2.942) and (1.371 ± 0.1127), respectively. The described techniques are designed to be simple and easy to implement with minimal training or expense, improving access to dense and/or metabolically active 3D cell culture models.

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Section: Image Acquisition Tissue Clearing and Volumetric Quantificationmentioning

confidence: 99%

In vitroformation and extended culture of highly metabolically active and contractile tissues

Bagdasarian

Tonmoy

Park

et al. 2023

Preprint

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“…Consequently, results are potentially less reproducible, incomplete, or misleading. For example, premature cell development and limited hierarchical structures in in vitro studies lead to morphological and functional deficiencies 19 – 21 , which can impact on the mechanical loading response of cells 9 . Generalisation to the broader clinical context is also limited 17 , 22 , hindering the real-world application of the research.…”

Section: Introductionmentioning

confidence: 99%

An ex vivo animal model to study the effect of transverse mechanical loading on skeletal muscle

Sargent,

Wark,

Day

et al. 2024

Commun Biol

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In many populations like wheelchair and prosthetic users, the soft tissue is subject to excessive or repetitive loading, making it prone to Deep Tissue Injury (DTI). To study the skeletal muscle response to physical stress, numerous in vitro and in vivo models exist. Yet, accuracy, variability, and ethical considerations pose significant trade-offs. Here, we present an ex vivo approach to address these limitations and offer additional quantitative information on cellular damage. In this study, skeletal muscle tissue from Sprague Dawley rats was isolated and transversely loaded. Histological analysis and fluorescence staining demonstrated that the setup was suitable to keep the tissue alive throughout the experimental procedure. Mechanically induced cell damage was readily distinguishable through morphological changes and uptake of a membrane impermeable dye. Our comparably simple experimental setup can be adapted to different loading conditions and tissues to assess the cell response to mechanical loading in future studies.

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“…For this, different procedures to engineer 3D in vivo‐like structures or bioartificial muscles (BAMs) were already described. [ 6–10 ] The most used materials as a biomimetic scaffold for BAMs are fibrin and collagen. [ 7,8,10–12 ] Gelatin, as a collagen derivate, is considered an interesting material for several reasons.…”

Section: Introductionmentioning

confidence: 99%

Ionically Modified Gelatin Hydrogels Maintain Murine Myogenic Cell Viability and Fusion Capacity

Burattini

Lippens

Baleine

et al. 2023

Macromolecular Bioscience

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For tissue engineering of skeletal muscles, there is a need for biomaterials which do not only allow cell attachment, proliferation, and differentiation, but also support the physiological conditions of the tissue. Next to the chemical nature and structure of the biomaterial, its response to the application of biophysical stimuli, such as mechanical deformation or application of electrical pulses, can impact in vitro tissue culture. In this study, gelatin methacryloyl (GelMA) is modified with hydrophilic 2‐acryloxyethyltrimethylammonium chloride (AETA) and 3‐sulfopropyl acrylate potassium (SPA) ionic comonomers to obtain a piezoionic hydrogel. Rheology, mass swelling, gel fraction, and mechanical characteristics are determined. The piezoionic properties of the SPA and AETA‐modified GelMA are confirmed by a significant increase in ionic conductivity and an electrical response as a function of mechanical stress. Murine myoblasts display a viability of >95% after 1 week on the piezoionic hydrogels, confirming their biocompatibility. The GelMA modifications do not influence the fusion capacity of the seeded myoblasts or myotube width after myotube formation. These results describe a novel functionalization providing new possibilities to exploit piezo‐effects in the tissue engineering field.

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Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

Cited by 4 publications

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In vitroformation and extended culture of highly metabolically active and contractile tissues

In vitroformation and extended culture of highly metabolically active and contractile tissues

An ex vivo animal model to study the effect of transverse mechanical loading on skeletal muscle

Ionically Modified Gelatin Hydrogels Maintain Murine Myogenic Cell Viability and Fusion Capacity

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