Bioinspired 3D Cocultures of Human Skeletal Myoblasts and Motoneuron-like Cells to Investigate Neuromuscular Function In Vitro
Dixon, Thomas.
2017
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Abstract: The physical connection in the developing embryo between ventral
horn motoneurons and skeletal muscle myoblasts is responsible for the creation of
neuromuscular junctions (NMJs), which allow electrical signals from the spinal cord to be
translated to mechanical work in contractile skeletal muscle. Pathology at the NMJ
contributes to the spectrum of neuromuscular, motoneuron, and ... read moredystrophic disease. Improving
in vitro tools which allow for recapitulation of the normal physiology of the neuromuscular
connection enables researchers to better understand the development and maturation of NMJs.
Further development of these tools and use of diseased cells will help to decipher
mechanisms leading to disruption of the NMJ, and related peripheral nerve and skeletal
muscle pathology. In this work, we describe a unique platform to coculture human skeletal
muscle myoblast-seeded hydrogels suspended in between flexible cantilevers with integrated
motoneuron-like cells derived from human induced neural stem cells. This platform is fully
customizable using 3D freeform printing into standard lab tissue culture materials, and
allows for quantifiable human myoblast alignment in 3D with precise motoneuron integration
into preformed myotubes. The coculture method allows for observation and analysis of
neurite outgrowth and myogenic differentiation in 3D with quantification of several
parameters of muscle innervation and function. We demonstrated activation of muscle
originated calcium transients in 3D constructs as well as some evidence of nerve originated
contractions in a fully human 3D NMJ. Future refinements with increased cell specificity
and external stimulation may lead to a high throughput method of neuromuscular analysis of
patient derived cells.
Thesis (Ph.D.)--Tufts University, 2017.
Submitted to the Dept. of Cell, Molecular & Developmental Biology.
Advisor: David Kaplan.
Keyword: Biomedical engineering.read less - ID:
- ww72bp65c
- Component ID:
- tufts:23389
- To Cite:
- TARC Citation Guide EndNote
