Field Neurosciences Institute Laboratory for Restorative Neurology, Central Michigan University, Mount Pleasant, MI 48859, USA; Program in Neurosciences, Central Michigan University, Mount Pleasant, MI 48859, USA.
Field Neurosciences Institute Laboratory for Restorative Neurology, Central Michigan University, Mount Pleasant, MI 48859, USA; Program in Neurosciences, Central Michigan University, Mount Pleasant, MI 48859, USA; Department of Psychology, Central Michigan University, Mount Pleasant, MI 48859, USA; College of Medicine, Central Michigan University, Mount Pleasant, MI 48859, USA.
Brain Res. 2018 Nov 15;1699:19-33. doi: 10.1016/j.brainres.2018.06.002. Epub 2018 Jun 5.
Transplanting stem cells engineered to overexpress trophic factors can improve motor abilities and facilitate axon regeneration following spinal cord injury. This study compared several transplantation paradigms using mesenchymal stem cells (MSCs) that overexpress the multi-neurotrophin, NT-3/D15A (NT-3-MSCs), to determine if different grafting strategies can elicit improved axon regeneration and/or behavioral outcomes following a complete T9 spinal transection. At one week post-transection, NT-3-MSCs were transplanted above, and at several locations below, the lesion site. A rostral-to-caudal gradient of NT-3-MSCs was produced by incrementally increasing the number of transplanted cells at locations distal to the transection. Motor function was analyzed using the Basso, Beattie, and Bresnahan scale for 7-weeks post-injury. The corticospinal tract was traced using biotinylated dextran amines, while raphespinal fibers were visualized using immunohistochemistry. Cell viability was assessed using transplants of NT-3-MSCs that express tdTomato. Retrograde tracing using fluorogold, as well as spinal re-transections, were performed to discriminate between a supra-spinal or reflexive influence of regained motor functions. NT-3-MSC transplants improved motor outcomes and tissue continuity at the transection site, however retrograde tracing using fluorogold revealed no evidence of axon regeneration. A spinal re-transection also failed to eliminate the improvement in motor outcomes produced by the transplant. We conclude that transplantation of NT-3-MSCs can improve motor function and morphological outcomes following a complete spinal transection without promoting axonal regeneration.
移植过表达神经营养因子的干细胞可以改善脊髓损伤后的运动能力并促进轴突再生。本研究比较了几种使用过表达多神经营养因子 NT-3/D15A(NT-3-MSCs)的间充质干细胞(MSCs)的移植范例,以确定不同的移植策略是否可以在完全 T9 脊髓横断后引起更好的轴突再生和/或行为结果。在横断后一周,将 NT-3-MSCs 移植到损伤部位上方和下方的多个部位。通过在远离横断部位的位置逐渐增加移植细胞的数量,产生了从头部到尾部的 NT-3-MSCs 梯度。在损伤后 7 周使用 Basso、Beattie 和 Bresnahan 量表分析运动功能。使用生物素化葡聚糖胺追踪皮质脊髓束,并用免疫组织化学观察中缝脊髓纤维。使用表达 tdTomato 的 NT-3-MSC 移植评估细胞活力。使用荧光金进行逆行追踪,以及对脊髓进行再横切,以区分恢复运动功能的是上运动神经元或反射性影响。NT-3-MSC 移植改善了运动结果和损伤部位的组织连续性,但使用荧光金进行逆行追踪没有发现轴突再生的证据。脊髓再横切也未能消除移植产生的运动结果改善。我们得出结论,移植 NT-3-MSCs 可以改善完全脊髓横断后的运动功能和形态结果,而不会促进轴突再生。
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