Home › Cell Biology › Investigating Functional Regeneration in Organotypic Spinal Cord Co-cultures Grown on Multi-electrode Arrays
Cell Biology JoVE (Open Access) Citable · DOI

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

DOI: 10.3791/53121-v
What you'll learn
  • ✓ Prepare and mount spinal cord tissue slices on multi-electrode arrays
  • ✓ Create mechanical lesions and record functional electrophysiological activity in vitro
  • ✓ Assess propriospinal regeneration and synchronized neural activity over time
Protocol

Biopharma Insights Here we present a protocol that is based on spinal cord slices cultured on multi-electrode arrays to study functional regeneration of propriospinal connections in vitro.

Difficulty
advanced
Total time
~3–4 days (tissue preparation, culture, lesion induction, and recording)
Model organism
Rat (spinal cord tissue)
Biosafety
BSL-1

Steps

1
Prepare multi-electrode arrays for culture

Clean, sterilize, and condition multi-electrode array devices for spinal cord tissue mounting. Ensure electrodes are properly functioning and coated as needed.

▶ 01:25
2
Dissect spinal cord tissue from model organism

Extract and isolate spinal cord tissue from the animal under sterile conditions, removing meninges and preparing tissue for sectioning.

▶ 02:39
3
Mount spinal cord slices on multi-electrode arrays

Position tissue slices onto the prepared electrode arrays and secure them with appropriate culture medium and supports to maintain contact with electrodes.

▶ 03:52
4
Create mechanical lesions in tissue slices

Perform controlled mechanical injury (transection or partial lesion) to the mounted spinal cord slices to model spinal cord damage and study regeneration.

▶ 05:09
5
Record spontaneous electrophysiological activity

Acquire multi-channel electrical recordings from the tissue to measure spontaneous neural activity and functional connectivity before and after lesion.

▶ 06:11
6
Analyze synchronized activity and age-related changes

Process recorded data to quantify synchronized burst activity and compare functional regeneration patterns across different tissue ages.

▶ 06:59
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