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In vitro stretch reflex arc
An In Vitro Model of Stem Cell Innervation of Myotubes
James J. Hickman: PI |
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| IDEA
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Random dissociated cell cultures have only a limited use in the study of complex physiological processes or diseases such as Amyotrophic Lateral Sclerosis (ALS) or spinal cord injury. Using surface chemistry and advanced patterning methods a functional model of the spinal stretch reflex arc can be created. This model will be an improvement over current in vitro models that are composed of disorganized culture systems because the interaction between the different cell types will be physiological ensuring a healthy development and in vivo - like functionality. The benefit of this system compared to in vivo models will be the accessibility of each element to experimental manipulations such as selective drug administration or replacement with cells from transgenic animals. |
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| GOAL |
Develop patterned artificial surfaces integrated with a microfabricated silicon device to create a physiologically realistic in vitro implementation of the stretch reflex arc in order to study normal and pathological behavior of this important functional unit of the spinal cord. |
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| APPROACH |
Skeletal muscle myotubes will be cultured on cantilevers and innervated by patterned motoneurons and DRG cells |
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| RESULTS |
We have developed serum-free culture conditions to culture motoneurons (Motoneuron
2004.pdf, Adult
spinal cord.pdf), skeletal muscle cells (Skeletal
Muscle.pdf) and DRG cells on the same MEMs device.
Embryonic motoneurons grown on patterns and embryonic skeletal muscle myotubes cultured in the same serum-free medium We have developed the methods to pattern myotubes using photolithography (C2C12.pdf)
Patterned C2C12 myotubes We have successfully demonstrated that skeletal muscle myotubes can be grown on cantilevers and contraction of the myotubes can be detected by the inflection of the cantilever
C2C12 myotubes on the cantilevers and measurement of contraction by the inflection of the cantilever We have started the mathematical analysis of the in vitro stretch reflex arc at the system-level (reflex arc.pdf) and at the component level by determining the transfer characteristics of each elements using white-noise analysis. We adapted this method to analyze the cell-electrode interface (Electrode.htm).
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| WORK IN PROGRESS | Enhance neuromuscular junction formation in
motoneuron-muscle co-cultures Enhance contractibility of skeletal muscle Pattern muscle and motoneurons on cantilevers Integrate DRG-cells in the co-cultures Development of the muscle spindle in vitro Development and integration of stem-cell derived motoneurons |
