Three-dimensional neuron–muscle constructs with neuromuscular junctions

Morimoto, Yuya; Kato-Negishi, Midori; Onoe, Hiroaki

doi:10.1016/j.biomaterials.2013.08.062

Cited by 170 publications

(183 citation statements)

References 31 publications

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“…In these studies, scaffold delivery of VEGF and IGF resulted in a 2.4-fold increase in peak tetanic force over blank scaffolds, whereas scaffold delivery of VEGF, IGF, and myoblasts resulted in 1.5-6.0-fold increases in peak tetanic force over control conditions (13,32,39). Although additional biomaterial-based approaches have induced similar improvements in muscle function through the codelivery of myogenic bioagents with endothelial cells to enhance vascularization (5.5-fold increase in active stress over blank scaffold) or neural stem cells to promote innervation of muscle constructs (2.0-fold increase in maximum tetanic force over nerve-deficient controls) (44)(45)(46)(47), no bioagents were delivered from the scaffolds in this study. It is possible that the magnetic field directly affected regeneration in the current study, as past studies involving treatment of myoblasts and myotubes with static magnetic fields have demonstrated increased myogenic differentiation and cell alignment (48,49).…”

Section: Markers Of Muscle Regeneration: Centrally Located Nuclei Andmentioning

confidence: 99%

Biologic-free mechanically induced muscle regeneration

Cezar

Roche

Vandenburgh

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

146

125

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Severe skeletal muscle injuries are common and can lead to extensive fibrosis, scarring, and loss of function. Clinically, no therapeutic intervention exists that allows for a full functional restoration. As a result, both drug and cellular therapies are being widely investigated for treatment of muscle injury. Because muscle is known to respond to mechanical loading, we investigated instead whether a material system capable of massage-like compressions could promote regeneration. Magnetic actuation of biphasic ferrogel scaffolds implanted at the site of muscle injury resulted in uniform cyclic compressions that led to reduced fibrous capsule formation around the implant, as well as reduced fibrosis and inflammation in the injured muscle. In contrast, no significant effect of ferrogel actuation on muscle vascularization or perfusion was found. Strikingly, ferrogel-driven mechanical compressions led to enhanced muscle regeneration and a ∼threefold increase in maximum contractile force of the treated muscle at 2 wk compared with no-treatment controls. Although this study focuses on the repair of severely injured skeletal muscle, magnetically stimulated bioagent-free ferrogels may find broad utility in the field of regenerative medicine.

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Section: Markers Of Muscle Regeneration: Centrally Located Nuclei Andmentioning

confidence: 99%

Biologic-free mechanically induced muscle regeneration

Cezar

Roche

Vandenburgh

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

146

125

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“…When we increased the glutamate concentration from 200 to 400 μM, the frequency of muscle twitching increased from 1.08 to 1.33 contractions per second, whereas the average displacement decreased from 6.3 ± 0 to 4.4 ± 0.2 μm per twitch (Figure 6e and Supplementary Video S4). The addition of 25 μM tubocurarine chloride (curare), an irreversible nicotinic NMJ antagonist and muscle relaxant 23 that blocks AChRs, halted the contractions; no further muscle twitching was observed.…”

Section: Chemical Stimulation Of Mns In Multi-layered Tissue Ringsmentioning

confidence: 99%

“…Beyond applications in regenerative medicine and therapeutics 20 , however, only a few studies have produced 3D NMJ platforms or applied neuromuscular research to living cellular systems using embryonic or neural stem cells [21][22][23] . Furthermore, there is a lack of research demonstrating the possibility of translating such an arrangement into a platform that is potentially autonomous, scalable, and forward-engineered, which are necessary characteristics of a mobile and functional cellular system or machine.…”

Section: Introductionmentioning

confidence: 99%

A 3D-printed platform for modular neuromuscular motor units

et al. 2017

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A complex and functional living cellular system requires the interaction of one or more cell types to perform specific tasks, such as sensing, processing, or force production. Modular and flexible platforms for fabrication of such multi-cellular modules and their characterization have been lacking. Here, we present a modular cellular system, made up of multi-layered tissue rings containing integrated skeletal muscle and motor neurons (MNs) embedded in an extracellular matrix. The MNs were differentiated from mouse embryonic stem cells through the formation of embryoid bodies (EBs), which are spherical aggregations of cells grown in a suspension culture. The EBs were integrated into a tissue ring with skeletal muscle, which was differentiated in parallel, to create a co-culture amenable to both cell types. The multi-layered rings were then sequentially placed on a stationary three-dimensionalprinted hydrogel structure resembling an anatomical muscle-tendon-bone organization. We demonstrate that the site-specific innervation of a group of muscle fibers in the multi-layered tissue rings allows for muscle contraction via chemical stimulation of MNs with glutamate, a major excitatory neurotransmitter in the mammalian nervous system, with the frequency of contraction increasing with glutamate concentration. The addition of tubocurarine chloride (a nicotinic receptor antagonist) halted the contractions, indicating that muscle contraction was MN induced. With a bio-fabricated system permitting controllable mechanical and geometric attributes in a range of length scales, our novel engineered cellular system can be utilized for easier integration of other modular "building blocks" in living cellular and biological machines.

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“…To overcome these problems, different in vitro coculture systems have been set up in which motor neuron and skeletal muscle are grown together in order to recapitulate the formation and eventual disruption of the NMJ. To date, co-culture methods established from various species have been described, including mouse (Morimoto et al, 2013;Zahavi et al, 2015), rat (Das et al, 2010;Southam et al, 2013), Xenopus (Lu et al, 1996;Peng et al, 2003) and chick (Frank and Fischbach, 1979), and also heterologous co-cultures built from motor neuron and muscle cells obtained from different species, such as rat-human , mouse-human (Son et al, 2011) and mouse-chick (Soundararajan et al, 2007). However, these co-culture methods resulted in the formation of immature myofibers (thin muscle fiber, with centrally localized nuclei and no transversal triads) with immature sarcomeric structures (Das et al, 2007(Das et al, , 2009Southam et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A system for studying mechanisms of neuromuscular junction development and maintenance

Vilmont¹,

Cadot²,

Ouanounou³

et al. 2016

Journal of Cell Science

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The neuromuscular junction (NMJ), a cellular synapse between a motor neuron and a skeletal muscle fiber, enables the translation of chemical cues into physical activity. The development of this special structure has been subject to numerous investigations, but its complexity renders in vivo studies particularly difficult to perform. In vitro modeling of the neuromuscular junction represents a powerful tool to delineate fully the fine tuning of events that lead to subcellular specialization at the pre-synaptic and post-synaptic sites. Here, we describe a novel heterologous co-culture in vitro method using rat spinal cord explants with dorsal root ganglia and murine primary myoblasts to study neuromuscular junctions. This system allows the formation and long-term survival of highly differentiated myofibers, motor neurons, supporting glial cells and functional neuromuscular junctions with post-synaptic specialization. Therefore, fundamental aspects of NMJ formation and maintenance can be studied using the described system, which can be adapted to model multiple NMJ-associated disorders.

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Three-dimensional neuron–muscle constructs with neuromuscular junctions

Cited by 170 publications

References 31 publications

Biologic-free mechanically induced muscle regeneration

Biologic-free mechanically induced muscle regeneration

A 3D-printed platform for modular neuromuscular motor units

A system for studying mechanisms of neuromuscular junction development and maintenance

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