Optically controlled contraction of photosensitive skeletal muscle cells

Asano, Takao; Ishizua, Toru; Yawo, Hiromu

doi:10.1002/bit.23285

Cited by 57 publications

(43 citation statements)

References 27 publications

(29 reference statements)

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“…Previous work showed that ChR2 enables temporally precise (tens of milliseconds) and spatially confined control of cardiac muscle contraction (17). A recent work reported the feasibility of optical control for skeletal muscle contractions (40). Here, we extend the results of this work by demonstrating selective activation of individual myotubes with confined illumination as well as formation, characterization and optical control of genetically engineered, 3D skeletal muscle microtissues.…”

Section: Resultsmentioning

confidence: 99%

Formation and optogenetic control of engineered 3D skeletal muscle bioactuators

et al. 2012

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Densely arrayed skeletal myotubes are activated individually and as a group using precise optical stimulation with high spatiotemporal resolution. Skeletal muscle myoblasts are genetically encoded to express light-activated cation channel, Channelrhodopsin-2, which allows for spatiotemporal coordination of the multitude of skeletal myotubes that contract in response to pulsed blue light. Furthermore, ensembles of mature functional 3D muscle microtissues have been formed from the optogenetically encoded myoblasts using a high-throughput device. The device, called “skeletal muscle on a chip”, not only provides the myoblasts with controlled stress and constraints necessary for muscle alignment, fusion and maturation, but also facilitates to measure forces and characterize the muscle tissue. We measured the specific static and dynamic stresses generated by the microtissues, and characterized the morphology and alignment of the myotubes within the constructs. The device allows for testing the effect of a wide range of parameters (cell source, matrix composition, microtissue geometry, auxotonic load, growth factors, and exercise) on the maturation, structure, and function of the engineered muscle tissues in a combinatorial manner. Our studies integrate tools from optogenetics and microelectromechanical systems (MEMS) technology with skeletal muscle tissue engineering to open up opportunities to generate soft robots actuated by multitude of spatiotemporally coordinated 3D skeletal muscle microtissues.

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Section: Resultsmentioning

confidence: 99%

Formation and optogenetic control of engineered 3D skeletal muscle bioactuators

et al. 2012

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“…For example, ChR2 has been expressed in neurons (19) to monitor and control their membrane potential, intracellular acidity, Ca 2+ influx, and other cellular processes (see ref (14) for review). In addition, ChR2 has been used to manipulate the membrane excitability of cardiomyocytes (20)(21)(22), skeletal muscle cells (23) and cell lines expressing voltage-gated ion channels (24,25).…”

Section: Introductionmentioning

confidence: 99%

Precisely control mitochondria with light to manipulate cell fate decision

Erñst

Yun

et al. 2018

Preprint

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Mitochondrial dysfunction has been implicated in many pathological conditions and diseases. The normal functioning of mitochondria relies on maintaining the inner mitochondrial membrane (IMM) potential (a.k.a. ΔΨ m ) that is essential for ATP synthesis, Ca 2+ homeostasis, redox balance and regulation of other key signaling pathways such as mitophagy and apoptosis.However, the detailed mechanisms by which ΔΨ m regulates cellular function remain incompletely understood, partially due to difficulty of manipulating ΔΨ m with spatiotemporal resolution, reversibility, or cell type specificity. To address this need, we have developed a nextgeneration optogenetic-based technique for controllable mitochondrial depolarization with light. We demonstrate successful targeting of the heterologous Channelrhodopsin-2 (ChR2) fusion protein to the IMM and formation of functional cationic channels capable of light-induced selective ΔΨ m depolarization and mitochondrial autophagy. Importantly, we for the first time show that optogenetic-mediated mitochondrial depolarization can be well-controlled to differentially influence the fate of cells expressing mitochondrial ChR2: while sustained moderate light illumination induces substantial apoptotic cell death, transient mild light illumination elicits cytoprotection via mitochondrial preconditioning. Finally, we show that Parkin overexpression exacerbates, instead of ameliorating, mitochondrial depolarization-mediated cell death in HeLa cells. In summary, we provide evidence that the described mitochondrial-targeted optogenetics may have a broad application for studying the role of mitochondria in regulating cell function and fate decision.

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“…Pulsatile electrical stimulation can also be used to induce and pace the muscle cells38. Recently, light-induced stimulation of genetically-engineered muscle cells that express the light-activated cation channels, halorhodopsin or channelrhodopsin-2 was demonstrated39404142. Integration of these engineered muscle cells to optically drive the bio-bots offers another exciting future possibility.…”

Section: Discussionmentioning

confidence: 99%

Development of Miniaturized Walking Biological Machines

Chan

Park

Collens

et al. 2012

Sci Rep

202

175

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The quest to ‘forward-engineer’ and fabricate biological machines remains a grand challenge. Towards this end, we have fabricated locomotive “bio-bots” from hydrogels and cardiomyocytes using a 3D printer. The multi-material bio-bot consisted of a ‘biological bimorph’ cantilever structure as the actuator to power the bio-bot, and a base structure to define the asymmetric shape for locomotion. The cantilever structure was seeded with a sheet of contractile cardiomyocytes. We evaluated the locomotive mechanisms of several designs of bio-bots by changing the cantilever thickness. The bio-bot that demonstrated the most efficient mechanism of locomotion maximized the use of contractile forces for overcoming friction of the supporting leg, while preventing backward movement of the actuating leg upon relaxation. The maximum recorded velocity of the bio-bot was ~236 µm s−1, with an average displacement per power stroke of ~354 µm and average beating frequency of ~1.5 Hz.

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Optically controlled contraction of photosensitive skeletal muscle cells

Cited by 57 publications

References 27 publications

Formation and optogenetic control of engineered 3D skeletal muscle bioactuators

Formation and optogenetic control of engineered 3D skeletal muscle bioactuators

Precisely control mitochondria with light to manipulate cell fate decision

Development of Miniaturized Walking Biological Machines

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