3D Timelapse Analysis of Muscle Satellite Cell Motility

Siegel, Ashley L.; Atchison, Kevin; Fisher, Kevin E.; Davis, George E.; Cornelison, D.D.W.

doi:10.1002/stem.178

Cited by 150 publications

(147 citation statements)

References 70 publications

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“…By culturing individual myofibers, satellite cell heterogeneity with respect to myofiber, muscle type or regenerative potential has been addressed 15 . Elaborate studies using live imaging of single cultured fibers allowed for the gaining information on satellite cell migration pattern 16 . The acquisition of quantitative data is also possible.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Pasut

Jones

Rudnicki

2013

JoVE

144

135

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Muscle regeneration in the adult is performed by resident stem cells called satellite cells. Satellite cells are defined by their position between the basal lamina and the sarcolemma of each myofiber. Current knowledge of their behavior heavily relies on the use of the single myofiber isolation protocol. In 1985, Bischoff described a protocol to isolate single live fibers from the Flexor Digitorum Brevis (FDB) of adult rats with the goal to create an in vitro system in which the physical association between the myofiber and its stem cells is preserved 1 . In 1995, Rosenblattmodified the Bischoff protocol such that myofibers are singly picked and handled separately after collagenase digestion instead of being isolated by gravity sedimentation 2,3 . The Rosenblatt or Bischoff protocol has since been adapted to different muscles, age or conditions [3][4][5][6] . The single myofiber isolation technique is an indispensable tool due its unique advantages. First, in the single myofiber protocol, satellite cells are maintained beneath the basal lamina. This is a unique feature of the protocol as other techniques such as Fluorescence Activated Cell Sorting require chemical and mechanical tissue dissociation 7 . Although the myofiber culture system cannot substitute for in vivo studies, it does offer an excellent platform to address relevant biological properties of muscle stem cells. Single myofibers can be cultured in standard plating conditions or in floating conditions. Satellite cells on floating myofibers are subjected to virtually no other influence than the myofiber environment. Substrate stiffness and coating have been shown to influence satellite cells' ability to regenerate muscles 8, 9 so being able to control each of these factors independently allows discrimination between niche-dependent and -independent responses. Different concentrations of serum have also been shown to have an effect on the transition from quiescence to activation. To preserve the quiescence state of its associated satellite cells, fibers should be kept in low serum medium [1][2][3] . This is particularly useful when studying genes involved in the quiescence state. In serum rich medium, satellite cells quickly activate, proliferate, migrate and differentiate, thus mimicking the in vivo regenerative process [1][2][3]

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Section: Discussionmentioning

confidence: 99%

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Pasut

Jones

Rudnicki

2013

JoVE

144

135

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“…Recently, we showed that activated primary satellite cells are highly motile in the context of their native substrate (the surface of the myofiber), moving up to 2.4 cm in 24 hours (Siegel et al, 2009). The same study found that activated satellite cells express mRNAs encoding multiple examples of classical guidance pathway ligands and receptors (Ephs, ephrins, semaphorins, plexins, neuropilins and Robos).…”

Section: Introductionmentioning

confidence: 94%

Eph/ephrin interactions modulate muscle satellite cell motility and patterning

et al. 2011

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SUMMARYDuring development and regeneration, directed migration of cells, including neural crest cells, endothelial cells, axonal growth cones and many types of adult stem cells, to specific areas distant from their origin is necessary for their function. We have recently shown that adult skeletal muscle stem cells (satellite cells), once activated by isolation or injury, are a highly motile population with the potential to respond to multiple guidance cues, based on their expression of classical guidance receptors. We show here that, in vivo, differentiated and regenerating myofibers dynamically express a subset of ephrin guidance ligands, as well as Eph receptors. This expression has previously only been examined in the context of muscle-nerve interactions; however, we propose that it might also play a role in satellite cell-mediated muscle repair. Therefore, we investigated whether Eph-ephrin signaling would produce changes in satellite cell directional motility. Using a classical ephrin 'stripe' assay, we found that satellite cells respond to a subset of ephrins with repulsive behavior in vitro; patterning of differentiating myotubes is also parallel to ephrin stripes. This behavior can be replicated in a heterologous in vivo system, the hindbrain of the developing quail, in which neural crest cells are directed in streams to the branchial arches and to the forelimb of the developing quail, where presumptive limb myoblasts emigrate from the somite. We hypothesize that guidance signaling might impact multiple steps in muscle regeneration, including escape from the niche, directed migration to sites of injury, cell-cell interactions among satellite cell progeny, and differentiation and patterning of regenerated muscle.

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“…However once they have emerged onto the muscle fiber, migration, one of the key activities undertaken by the stem cells is severely limited by the absence of glycolytic activity [7,20]. Our results show that this process must be highly energy dependent since the cells unable to fully undertake glycolysis decrease their migration rate by over 50%.…”

Section: Discussionmentioning

confidence: 85%

Investigating the Influence of Extracellular Matrix and Glycolytic Metabolism on Muscle Stem Cell Migration on Their Native Fiber Environment

Butera

Collins‐Hooper

Mitchell

et al. 2015

Fibers

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Abstract:The composition of the extracellular matrix (ECM) of skeletal muscle fibers is a unique environment that supports the regenerative capacity of satellite cells; the resident stem cell population. The impact of environment has great bearing on key properties permitting satellite cells to carry out tissue repair. In this study, we have investigated the influence of the ECM and glycolytic metabolism on satellite cell emergence and migration-two early processes required for muscle repair. Our results show that both influence the rate at which satellite cells emerge from the sub-basal lamina position and their rate of migration. These studies highlight the necessity of performing analysis of satellite behavior on their native substrate and will inform on the production of artificial scaffolds intended for medical uses.

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3D Timelapse Analysis of Muscle Satellite Cell Motility

Cited by 150 publications

References 70 publications

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Eph/ephrin interactions modulate muscle satellite cell motility and patterning

Investigating the Influence of Extracellular Matrix and Glycolytic Metabolism on Muscle Stem Cell Migration on Their Native Fiber Environment

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