Three-dimensional contractile muscle tissue consisting of human skeletal myocyte cell line

Shima, Ai; Morimoto, Yuya; Sweeney, H. Lee; Takeuchi, Shoji

doi:10.1016/j.yexcr.2018.06.015

Cited by 30 publications

(20 citation statements)

References 23 publications

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“…In addition, engineered 3D skeletal muscle models mimic native muscle architecture 21 , provide structural integrity for long-term culture of myotubes in vitro, and enable contractile force measurements 22 . Recent articles report successful development of 3D culture models of human skeletal muscle 9,[23][24][25][26][27][28][29][30][31][32][33] . In these studies, active force is quantified on tissues removed from the supporting culture device to implement a force transducer, which is precise, but invasive.…”

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confidence: 99%

A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength

Afshar

Abraha

Bakooshli

et al. 2020

Sci Rep

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Three-dimensional (3D) in vitro models of human skeletal muscle mimic aspects of native tissue structure and function, thereby providing a promising system for disease modeling, drug discovery or pre-clinical validation, and toxicity testing. Widespread adoption of this research approach is hindered by the lack of easy-to-use platforms that are simple to fabricate and that yield arrays of human skeletal muscle micro-tissues (hMMTs) in culture with reproducible physiological responses that can be assayed non-invasively. Here, we describe a design and methods to generate a reusable mold to fabricate a 96-well platform, referred to as MyoTACTIC, that enables bulk production of 3D hMMTs. All 96-wells and all well features are cast in a single step from the reusable mold. Non-invasive calcium transient and contractile force measurements are performed on hMMTs directly in MyoTACTIC, and unbiased force analysis occurs by a custom automated algorithm, allowing for longitudinal studies of function. Characterizations of MyoTACTIC and resulting hMMTs confirms the capability of the device to support formation of hMMTs that recapitulate biological responses. We show that hMMT contractile force mirrors expected responses to compounds shown by others to decrease (dexamethasone, cerivastatin) or increase (IGF-1) skeletal muscle strength. Since MyoTACTIC supports hMMT long-term culture, we evaluated direct influences of pancreatic cancer chemotherapeutics agents on contraction competent human skeletal muscle myotubes. A single application of a clinically relevant dose of Irinotecan decreased hMMT contractile force generation, while clear effects on myotube atrophy were observed histologically only at a higher dose. This suggests an off-target effect that may contribute to cancer associated muscle wasting, and highlights the value of the MyoTACTIC platform to non-invasively predict modulators of human skeletal muscle function. Skeletal muscle is one of the most abundant tissues in the human body and it enables critical physiological and functional activities, such as thermogenesis 1 and mobility 2. There are many degenerative and fatal diseases of skeletal muscle that remain untreated and the underlying pathology of some muscle related diseases is not fully understood. The use of animal models to study skeletal muscle diseases has improved our understanding of in vivo drug response and disease pathology 3,4. However, in some cases animal models fail to accurately predict drug response in humans, in part due to species specific differences leading to inaccurate disease symptoms 5,6. Furthermore, animal models are expensive and time consuming making them less desirable for drug testing 7. As a result, a push to establish in vitro models of human skeletal muscle with reliable phenotypic readouts for drug testing is underway with the goal of improving therapeutic outcomes in humans.

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mentioning

confidence: 99%

A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength

Afshar

Abraha

Bakooshli

et al. 2020

Sci Rep

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“…However, it is also quite possible that 3D human myofiber culture systems require additional modifications to allow cores, and other features of CCD, to form in a dish. Although 3D cultures are enriched for larger-diameter myofibers than those observed in conventional 2D cultures at similar time points Shima et al, 2018), in vitro myofibers are still smaller than adult human skeletal-muscle ones. Thus, several strategies are being pursued to boost 3D-cultured human myotube maturation.…”

Section: D Human Cell Culture Modelsmentioning

confidence: 98%

“…Myotubes that form in engineered 3D human skeletal-muscle tissue systems are structurally more mature than those in conventional 2D culture, and do not need specialized culture supplements (Falcone et al, 2014;Guo et al, 2014;Mills et al, 2019;Pimentel et al, 2017). Specifically, 3D-cultured myofibers express higher levels of adult forms of myosin heavy chains , they express SERCA2 and functional RYRs (Madden et al, 2015;Shima et al, 2018), can undergo repeated rounds of contraction in response to electrical and chemical stimulation, and exhibit in vivo-like Ca 2+ handling (Madden et al, 2015).…”

Section: D Human Cell Culture Modelsmentioning

confidence: 99%

“…Interestingly, amorphous central cores that lack enzymatic activity, the defining feature of core myopathies, are most often observed in type 1 (slow twitch) myofibers (Sato et al, 2008;Wu et al, 2006). The premature loss of the most mature myotubes in conventional 2D culture impedes the study of mature fiber types, and this can partially be overcome by 3D modeling (Shima et al, 2018). The development of a human model in which different fiber types develop would give insight into why these myofibers are more prone to core lesions.…”

Section: D Human Cell Culture Modelsmentioning

confidence: 99%

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Cored in the act: the use of models to understand core myopathies

Fusto

Moyle

Gilbert

et al. 2019

Disease Models &Amp; Mechanisms

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The core myopathies are a group of congenital myopathies with variable clinical expressionranging from early-onset skeletal-muscle weakness to later-onset disease of variable severitythat are identified by characteristic 'core-like' lesions in myofibers and the presence of hypothonia and slowly or rather non-progressive muscle weakness. The genetic causes are diverse; central core disease is most often caused by mutations in ryanodine receptor 1 (RYR1), whereas multi-minicore disease is linked to pathogenic variants of several genes, including selenoprotein N (SELENON), RYR1 and titin (TTN). Understanding the mechanisms that drive core development and muscle weakness remains challenging due to the diversity of the excitation-contraction coupling (ECC) proteins involved and the differential effects of mutations across proteins. Because of this, the use of representative models expressing a mature ECC apparatus is crucial. Animal models have facilitated the identification of disease progression mechanisms for some mutations and have provided evidence to help explain genotype-phenotype correlations. However, many unanswered questions remain about the common and divergent pathological mechanisms that drive disease progression, and these mechanisms need to be understood in order to identify therapeutic targets. Several new transgenic animals have been described recently, expanding the spectrum of core myopathy models, including mice with patient-specific mutations. Furthermore, recent developments in 3D tissue engineering are expected to enable the study of core myopathy disease progression and the effects of potential therapeutic interventions in the context of human cells. In this Review, we summarize the current landscape of core myopathy models, and assess the hurdles and opportunities of future modeling strategies.

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“…In contrast, modern reconstituted 3D in vitro skeletal muscle systems are demonstrated to be an efficient tool for rapid and reliable drug screening [2,54] and allow for testing of personalized treatments on cells harvested from individual patients. Besides these medical advantages, such functional muscle tissues were reported to successfully mimic native muscle tissue with long-term structural integrity [23,27] and allow new insights and fundamental knowledge of muscle tissue development, force generation during contraction as well as the phases of disease onset and progression [2,11,19,24,25,31,32,41,48,51]. In culture platforms allowing for in situ force measurements, 3D skeletal muscle tissues self organize between mm sized posts around which muscle precursor cells are seeded together with an extracellular matrix.…”

Section: Introductionmentioning

confidence: 99%

Global and local tension measurements in biomimetic skeletal muscle tissues reveals early mechanical homeostasis

Hofemeier

Limon

Muenker

et al. 2020

Preprint

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AbstractThe mechanical properties and tension of muscle tissue are tightly related to proper skeletal muscle function, which makes experimental access to the biomechanics of muscle tissue development a key requirement to advance our understanding of muscle function and development. Recently developed elastic in vitro culture chambers allow for raising 3D muscle tissue under controlled conditions and measurements of tissue force generation. However, these chambers are inherently incompatible with high resolution microscopy limiting their usability to global force measurements, and preventing the exploitation of modern fluorescence based investigation methods for live and dynamic measurements. Here we present a new chamber design pairing global force measurements, quantified from post deflection, with local tension measurements obtained from elastic hydrogel beads embedded in the muscle tissue. High resolution 3D video microscopy of engineered muscle development, enabled by the new chamber, shows an early mechanical tissue homeostasis that remains stable in spite of continued myotube maturation.

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Three-dimensional contractile muscle tissue consisting of human skeletal myocyte cell line

Cited by 30 publications

References 23 publications

A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength

A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength

Cored in the act: the use of models to understand core myopathies

Global and local tension measurements in biomimetic skeletal muscle tissues reveals early mechanical homeostasis

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