In Vitro Tissue‐Engineered Skeletal Muscle Models for Studying Muscle Physiology and Disease

Prabhu, Neel K.; Wang, Jason; Bursac, Nenad

doi:10.1002/adhm.201701498

Cited by 86 publications

(82 citation statements)

References 279 publications

(334 reference statements)

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“…With respect to matrix-free 3D cultures, spheroids are common due to their ease and reliability of production. Currently, numerous 3D-spheroid models for tissues like skin and its pathological conditions (Chiricozzi et al, 2017;Klicks et al, 2019), tumor (Shroyer, 2016), intestine (Pereira et al, 2016), skeletal muscle (Khodabukus et al, 2018), or brain (Lee et al, 2017) are available.…”

Section: Introductionmentioning

confidence: 99%

Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward

Nürnberg

Vitacolonna

Klicks

et al. 2020

Front. Mol. Biosci.

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Three-dimensional cell cultures, such as spheroids and organoids, serve as increasingly important models in fundamental and applied research and start to be used for drug screening purposes. Optical tissue clearing procedures are employed to enhance visualization of fluorescence-stained organs, tissues, and three-dimensional cell cultures. To get a more systematic overview about the effects and applicability of optical tissue clearing on three-dimensional cell cultures, we compared six different clearing/embedding protocols on seven types of spheroid-and chip-based threedimensional cell cultures of approximately 300 µm in size that were stained with nuclear dyes, immunofluorescence, cell trackers, and cyan fluorescent protein. Subsequent whole mount confocal microscopy and semi-automated image analysis were performed to quantify the effects. Quantitative analysis included fluorescence signal intensity and signal-to-noise ratio as a function of z-depth as well as segmentation and counting of nuclei and immunopositive cells. In general, these analyses revealed five key points, which largely confirmed current knowledge and were quantified in this study. First, there was a massive variability of effects of different clearing protocols on sample transparency and shrinkage as well as on dye quenching. Second, all tested clearing protocols worked more efficiently on samples prepared with one cell type than on co-cultures. Third, z-compensation was imperative to minimize variations in signal-tonoise ratio. Fourth, a combination of sample-inherent cell density, sample shrinkage, uniformity of signal-to-noise ratio, and image resolution had a strong impact on data segmentation, cell counts, and relative numbers of immunofluorescence-positive cells. Finally, considering all mentioned aspects and including a wish for simplicity and speed of protocols-in particular, for screening purposes-clearing with 88% Glycerol appeared to be the most promising option amongst the ones tested.

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Section: Introductionmentioning

confidence: 99%

Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward

Nürnberg

Vitacolonna

Klicks

et al. 2020

Front. Mol. Biosci.

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“…Other cells such as chondrocytes rapidly lose their molecular signature and quickly dedifferentiate when removed from the joint environment [ 17 , 18 ]. Moreover, primary muscle cells are very sensitive to their physical environment; therefore, these cells are prone to detaching and limiting their mature phenotype on stiff substrates [ 19 ]. Additionally, relevant human tissue or cell samples are often difficult to obtain, sometimes requiring invasive surgery or only becoming available post-mortem [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Versatility of Induced Pluripotent Stem Cells (iPSCs) for Improving the Knowledge on Musculoskeletal Diseases

Sanjurjo‐Rodríguez

Castro-Viñuelas

Piñeiro-Ramil

et al. 2020

IJMS

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Induced pluripotent stem cells (iPSCs) represent an unlimited source of pluripotent cells capable of differentiating into any cell type of the body. Several studies have demonstrated the valuable use of iPSCs as a tool for studying the molecular and cellular mechanisms underlying disorders affecting bone, cartilage and muscle, as well as their potential for tissue repair. Musculoskeletal diseases are one of the major causes of disability worldwide and impose an important socio-economic burden. To date there is neither cure nor proven approach for effectively treating most of these conditions and therefore new strategies involving the use of cells have been increasingly investigated in the recent years. Nevertheless, some limitations related to the safety and differentiation protocols among others remain, which humpers the translational application of these strategies. Nonetheless, the potential is indisputable and iPSCs are likely to be a source of different types of cells useful in the musculoskeletal field, for either disease modeling or regenerative medicine. In this review, we aim to illustrate the great potential of iPSCs by summarizing and discussing the in vitro tissue regeneration preclinical studies that have been carried out in the musculoskeletal field by using iPSCs.

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“…Functional in vitro models of skeletal muscle would greatly benefit from the development of these advanced compound screening systems for drug efficacy/ toxicity studies. In these models, compounds can be tested for their impact on pathways that affect functional parameters such as muscle strength and fatigability in normal and diseased tissues (Khodabukus, Prabhu, Wang, & Bursac, 2018;A. S. T. Smith, Davis, Lee, Mack, & Kim, 2016;Vandenburgh, 2010).…”

Section: Introductionmentioning

confidence: 99%

A multiplexed in vitro assay system for evaluating human skeletal muscle functionality in response to drug treatment

Najjar

Smith

Long

et al. 2019

Biotech & Bioengineering

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In vitro systems that mimic organ functionality have become increasingly important tools in drug development studies. Systems that measure the functional properties of skeletal muscle are beneficial to compound screening studies and also for integration into multiorgan devices. To date, no studies have investigated human skeletal muscle responses to drug treatments at the single myotube level in vitro. This report details a microscale cantilever chip-based assay system for culturing individual human myotubes. The cantilevers, along with a laser and photo-detector system, enable measurement of myotube contractions in response to broad-field electrical stimulation. This system was used to obtain baseline functional parameters for untreated human myotubes, including peak contractile force and time-to-fatigue data. The cultured myotubes were then treated with known myotoxic compounds and the resulting functional changes were compared to baseline measurements as well as known physiological responses in vivo. The collected data demonstrate the system's capacity for screening direct effects of compound action on individual human skeletal myotubes in a reliable, reproducible, and noninvasive manner. Furthermore, it has the potential to be utilized for high-content screening, disease modeling, and exercise studies of human skeletal muscle performance utilizing iPSCs derived from specific patient populations such as the muscular dystrophies. K E Y W O R D Sbody-on-a-chip, cantilevers, functional assay, human, skeletal muscle

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In Vitro Tissue‐Engineered Skeletal Muscle Models for Studying Muscle Physiology and Disease

Cited by 86 publications

References 279 publications

Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward

Routine Optical Clearing of 3D-Cell Cultures: Simplicity Forward

Versatility of Induced Pluripotent Stem Cells (iPSCs) for Improving the Knowledge on Musculoskeletal Diseases

A multiplexed in vitro assay system for evaluating human skeletal muscle functionality in response to drug treatment

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