Home › Cell Biology › Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus
Cell Biology JoVE (Open Access) Citable · DOI

Surgical Transplantation of Mouse Neural Stem Cells into the Spinal Cords of Mice Infected with Neurotropic Mouse Hepatitis Virus

DOI: 10.3791/2834-v
What you'll learn
  • ✓ Perform thoracic laminectomy at T9 vertebra in mice
  • ✓ Prepare and inject neural stem cells into spinal cord
  • ✓ Manage pre- and post-operative care for transplanted mice
  • ✓ Assess successful NSC engraftment in demyelinated spinal tissue
Protocol

The transplantation of mouse neural stem cells (NSCs) into the spinal cords of mice with established demyelination is detailed. The preparation of NSCs, the laminectomy of thoracic vertebra 9 (T9), and transplantation of NSCs is outlined along with the pre- and post-operative care of the mice.

Difficulty
advanced
Total time
~2–3 hours per mouse (surgery + recovery monitoring)
Model organism
Mouse (C57BL/6J or equivalent, infected with neurotropic MHV)
Biosafety
BSL-2

Steps

1
Prepare neural stem cells for transplantation

Culture and prepare mouse NSCs to appropriate concentration and viability for surgical injection. Ensure cells are ready immediately before transplantation.

▶ 01:25
2
Anesthetize mouse and expose spinal cord

Induce general anesthesia, position mouse prone, and perform laminectomy at thoracic vertebra 9 (T9) to expose the dorsal spinal cord surface.

▶ 02:54
3
Inject neural stem cells into spinal cord

Using a Hamilton syringe or micropipette, carefully inject NSC suspension into the demyelinated spinal cord tissue at the T9 level with precise depth control.

▶ 04:45
4
Close surgical site and manage post-operative recovery

Suture muscle and skin layers, monitor anesthesia reversal, and provide analgesia and supportive care during immediate post-operative period.

▶ 07:00
5
Verify successful NSC transplantation

Assess graft integration and engraftment using histology or imaging at defined post-operative timepoints to confirm NSC survival and migration.

▶ 07:53
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