Lee Yee-Shuan, Wu Siliang, Arinzeh Treena Livingston, Bunge Mary Bartlett
The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine.
Department of Materials Science and Engineering, New Jersey Institute of Technology.
J Vis Exp. 2017 Nov 3(129):56077. doi: 10.3791/56077.
Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord injury. The complete transection model, although not as clinically relevant as the contusion model, is the most rigorous method to evaluate axon regeneration. In the contusion model, it is difficult to distinguish regenerated from sprouted or spared axons due to the presence of remaining tissue post injury. In the complete transection model, a bridging method is necessary to fill the gap and create continuity from the rostral to the caudal stumps in order to evaluate the effectiveness of the treatments. A reliable bridging surgery is essential to test outcome measures by reducing the variability due to the surgical method. The protocols described here are used to prepare Schwann cells (SCs) and conduits prior to transplantation, complete transection of the spinal cord at thoracic level 8 (T8), insert the conduit, and transplant SCs into the conduit. This approach also uses in situ gelling of an injectable basement membrane matrix with SC transplantation that allows improved axon growth across the rostral and caudal interfaces with the host tissue.
在大鼠脊髓损伤的各种模型中,挫伤模型是最常用的,因为它是人类脊髓损伤最常见的类型。完全横断模型虽然在临床相关性上不如挫伤模型,但却是评估轴突再生最严格的方法。在挫伤模型中,由于损伤后仍有残留组织,很难区分再生轴突与发芽或未受损的轴突。在完全横断模型中,需要一种桥接方法来填补间隙并建立从近端到远端残端的连续性,以便评估治疗效果。可靠的桥接手术对于通过减少手术方法导致的变异性来测试结果指标至关重要。这里描述的方案用于在移植前制备雪旺细胞(SCs)和导管,在胸8(T8)水平完全横断脊髓,插入导管,并将SCs移植到导管中。这种方法还使用了可注射基底膜基质与SCs移植的原位凝胶化,这允许改善轴突穿过与宿主组织的近端和远端界面的生长。