Compositions and methods for treatment of spinal cord injury
A technique for spinal cord injury, composition, applied in the field of compositions and methods for treating spinal cord injury, capable of addressing lack of success, no recovery of neurological function, no evidence of growth/extension of neurons and/or axons across the injury site, etc. question
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Embodiment 1
[0091] Materials and methods
[0092] Overview of Materials and Methods
[0093] Animal model: adult female Sprague-Dawley rats. Injury Model: Mid neck (C3-4) unilateral contusion. Treatment: Injection of EC alone, or in combination with NPC. Duration of treatment: Single delivery 1 week after injury. Therapeutic dose: 10 microliters containing 100,000 cells / ul culture medium (HBSS). Administration route: Inject directly into the affected area. Delivery method: Injection via a surgical syringe (Hamilton) with a 30 gauge steel needle. Experimental time: 7 weeks after injury. Anatomical Outcome Determination: Neuroanatomical Tracing and Immunohistochemistry (observation of effects on lesion cavity, vessels and axon growth). Functional Outcome Measures: Terminal Electrophysiology (observation of effects on muscle activity). Behavioral Outcome Measures: Weekly plethysmographic assessment (observe the effect on breathing patterns (respiratory rate, respiratory volume, m...
Embodiment 2
[0105] Effects of NPC and E4ORF1+EC transplantation in spinal cord injury model
[0106] Figure 1 provides a schematic representation of the methods and timeline used in the experiments described. Figure 1A. Neural progenitor cells (NPCs) were isolated from the developing rat spinal cord, cultured, frozen and thawed 1 day before transplantation. Figure 1B, Mouse spinal cord endothelial cells (EC) expressing E4ORF1 were thawed and cultured with lenti-GFP virus before transplantation. Figure 1C, 1 week after spinal cord contusion, NPCs and ECs were transplanted into the center of the lesion at a ratio of 1:1 (1,000,000 cells in total). Efficacy of this transplantation model was evaluated using a battery of anatomical (anterograde and retrograde tracers) and functional (terminal septal electromyography, dEMG) assessments. The experimental timeline is shown in Figure 1D.
[0107] Phenotypic analysis of transplanted NPCs and ECs showed differentiation into GFAP-positive glial ...
Embodiment 3
[0113] The role of glial progenitor cells or glial cells combined with E4ORF1+EC transplantation in spinal cord injury models
[0114] As described in Example 2, we found that after NPC transplantation, NPC differentiated into GFAP-positive glial cells about 6 weeks after transplantation. Therefore, we hypothesized that if glial progenitor cells or glial cells (instead of NPCs) were transplanted together with E4ORF1+ ECs, the above SCI repair could also be achieved.
[0115] To test this hypothesis, the following experiments were performed and all methods were as described above unless otherwise stated.
[0116] Glial progenitor cells and / or glial cells are obtained. Spinal cord endothelial cells (EC) were obtained and transformed to produce E4ORF1+EC as described above. The first combination of glial progenitor cells and E4ORF1+ECs and the second combination of glial cells and E4ORF1+ECs were transplanted into the center of the lesion in the above SCI model. Efficacy of ...
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