A 3D model of human skeletal muscle innervated with stem cell-derived motor neurons enables epsilon-subunit targeted myasthenic syndrome studies
Posted on: 12 March 2018
Preprint posted on 3 March 2018
A novel 3D co-culture system has been developed that can recapitulate human adult neuromuscular junctions in vitro
Selected by Chris Demers
Categories: bioengineering, cell biology, developmental biology
Background: Neuromuscular diseases are complex and devastating diseases, which in many cases only become symptomatic later in adult lives. Scientists have thus far failed to develop a biologically relevant model of adult human neuromuscular junctions, a critical site of disease pathogenesis. Such a platform would enable the study of not only the fundamental biology underpinning these diseases, but also provide a platform to test potential therapies.
Key Findings: This preprint represents the first example of an in vitro culture system that can be used to study adult human neuromuscular junctions. Critically, the authors found that the system only worked when cells were grown in 3D culture, as their 2D counterparts failed to undergo the critical developmental switch in acetylcholine receptors that is a hallmark of mature adult motor neurons.
Skeletal muscle precursors and stem cell derived motor neuron precursors were mixed in a fibrin/Geltrex™ hydrogel and placed in a PDMS microchannel with Velcro™ anchors for support. Over the course of just two-weeks the two populations self-organized and formed functional neuromuscular junctions. These junctions demonstrated many of the hallmarks of motor neuron innervated muscles, including localised calcium transients upon stimulation with glutamate.
Why I Chose This: I find it remarkable that stem cells have an inherent ability to self-organize into physiologically relevant structures. In most cases all that is required are the appropriate boundary conditions, be it soluble (i.e. addition of morphogens) or physical. In this preprint, by constraining cells to a three-dimensional microenvironment the authors were able to guide cells into forming functional, innervated muscles.
Open Questions:
- What are the unique aspects of the 3D culture conditions that lead to the maturation of the motor neurons?
- Is cell density a critical parameter to obtain reproducible results?
- Can the system be used with other motor neuron subtypes (i.e. MMC vs. LMC) and/or other muscle subtypes?
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