Investigating Functional Regeneration in Organotypic Spinal Cord Co-cultures Grown on Multi-electrode Arrays

Journal of Visualized Experiments, 2015 · DOI: 10.3791/53121 · Published: September 23, 2015

Simple Explanation

This study introduces an in vitro model using two organotypic spinal cord sections from embryonic rats, cultured on multi-electrode arrays (MEAs). These slices fuse together, allowing researchers to create lesions and study functional regeneration. The MEAs enable non-invasive, high-resolution recording of neuronal activity, while the organotypic slices maintain the original tissue architecture and local synaptic circuitry. This combination provides a sophisticated way to investigate intraspinal connections in isolation. The model allows researchers to assess functional connection of the slices by calculating the amount of synchronized bursts between the two sides and to analyze the slices morphologically by performing immunohistochemical stainings after the recordings.

Study Duration

8 to 28 DIV

Participants

E14 rat embryos

Evidence Level

Not specified

Key Findings

1
Cultures lesioned at a young age (7 - 9 DIV) displayed a high amount of synchronized activity 2-3 weeks after lesion, indicating a greater capacity for regeneration.
2
Cultures lesioned at later stages (>19 DIV) showed a distinct reduction in the ability to regenerate, suggesting that the regeneration ability decreases as the cultures mature.
3
Experiments using the nicotinic antagonist Mecamylamine (MEC) suggest that cholinergic synapses do not significantly contribute to the functional connection between slices in this model.

Research Summary

This study presents an in vitro model using organotypic spinal cord co-cultures on MEAs to investigate functional regeneration after spinal cord injury. The model allows for the study of propriospinal connections without interference from ascending and descending fiber tracts. Key findings indicate that the regeneration ability of functional connections decreases with culture age and that cholinergic synapses do not significantly contribute to functional connections between slices. The authors emphasize that the model can help to obtain insight about functional regeneration in the spinal cord, and provides stable conditions to investigate synaptic formation and synaptic plasticity in isolation and in great detail.

Practical Implications

Drug Discovery

The model could be used to screen potential therapeutic compounds aimed at promoting functional regeneration after spinal cord injury.

Understanding Spinal Cord Repair

The in vitro model provides a simplified system to dissect the mechanisms underlying spinal cord regeneration, focusing on propriospinal connections.

Optimizing Intervention Timing

The finding that regeneration capacity decreases with age suggests that interventions may be more effective if applied earlier after injury.

Study Limitations

1
The model lacks supraspinal and sensory input, limiting its ability to fully replicate in vivo conditions.
2
The culture preparation process represents a situation of CNS damage, potentially affecting glial cell behavior and tissue organization.
3
There is no evidence for the involvement of myelin associated inhibitors, e.g., Nogo-A, in the low regeneration potential of cultures lesioned at an old age.

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