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Protocol Steps
View Abstract
Spinal cord injury (SCI) results in loss of oligodendrocytes demyelination of surviving axons and severe functional impairment. Spontaneous remyelination is limited. Thus, cell replacement therapy is an attractive approach for myelin repair. In this study, we transplanted adult brain-derived neural precursor cells (NPCs) isolated from yellow fluorescent protein-expressing transgenic mice into the injured spinal cord of adult rats at 2 and 8 weeks after injury, which represents the subacute and chronic phases of SCI. A combination of growth factors, the anti-inflammatory drug minocycline, and cyclosporine A immunosuppression was used to enhance the survival of transplanted adult NPCs. Our results show the presence of a substantial number of surviving NPCs in the injured spinal cord up to 10 weeks after transplantation at the subacute stage of SCI. In contrast, cell survival was poor after transplantation into chronic lesions. After subacute transplantation, grafted cells migrated >5 mm rostrally and caudally. The surviving NPCs integrated principally along white-matter tracts and displayed close contact with the host axons and glial cells. Approximately 50% of grafted cells formed either oligodendroglial precursor cells or mature oligodendrocytes. NPC-derived oligodendrocytes expressed myelin basic protein and ensheathed the axons. We also observed that injured rats receiving NPC transplants had improved functional recovery as assessed by the Basso, Beattie, and Bresnahan Locomotor Rating Scale and grid-walk and footprint analyses. Our data provide strong evidence in support of the feasibility of adult NPCs for cell-based remyelination after SCI.
1
Spinal cord injury induction
Adult rats received spinal cord injury
not specifiednot specified
Note: Injury was performed prior to NPC transplantation
View evidence from paper
“transplanted adult brain-derived neural precursor cells (NPCs) isolated from yellow fluorescent protein-expressing transgenic mice into the injured spinal cord of adult rats”
2
NPC transplantation - subacute phase
Transplant adult NPCs into injured spinal cord at 2 weeks after injury (subacute phase)
2 weeks post-injurynot specified
Note: Represents subacute phase of spinal cord injury
View evidence from paper
“transplanted adult brain-derived neural precursor cells (NPCs) into the injured spinal cord of adult rats at 2 and 8 weeks after injury, which represents the subacute and chronic phases of SCI”
3
NPC transplantation - chronic phase
Transplant adult NPCs into injured spinal cord at 8 weeks after injury (chronic phase)
8 weeks post-injurynot specified
Note: Represents chronic phase of spinal cord injury
View evidence from paper
“transplanted adult brain-derived neural precursor cells (NPCs) into the injured spinal cord of adult rats at 2 and 8 weeks after injury, which represents the subacute and chronic phases of SCI”
4
Administer supportive pharmacological treatment
Administer combination of growth factors, minocycline, and cyclosporine A to enhance NPC survival
not specifiednot specified
Note: Treatment protocol to improve transplanted cell survival
View evidence from paper
“A combination of growth factors, the anti-inflammatory drug minocycline, and cyclosporine A immunosuppression was used to enhance the survival of transplanted adult NPCs”
5
Footprint analysis assessment
Perform footprint analysis to assess functional neurological recovery and locomotor function in injured rats receiving NPC transplants
Note: Conducted alongside Basso, Beattie, and Bresnahan Locomotor Rating Scale and grid-walk analyses
View evidence from paper
“injured rats receiving NPC transplants had improved functional recovery as assessed by the Basso, Beattie, and Bresnahan Locomotor Rating Scale and grid-walk and footprint analyses”
Subjects / Specimens
Species
rat
Strain
adult rats
Age
adult
Sex
unknown
Weight
not specified
Rats received spinal cord injury and NPC transplantation at 2 and 8 weeks after injury
