Wright Karina T, Uchida Kenzo, Bara Jennifer J, Roberts Sally, El Masri Wagih, Johnson William E B
Centre for Spinal Studies, Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry, Shropshire SY10 7AG, UK; Institute for Science and Technology in Medicine, Keele University, Keele, Staffordshire ST5 5BG, UK.
Department of Orthopaedics and Rehabilitation Medicine, University of Fukuimore, 3 - 9 - 1 Bunkyo, Fukui, Fukui Prefecture 910-8507, Japan.
Spine J. 2014 Aug 1;14(8):1722-33. doi: 10.1016/j.spinee.2014.01.021. Epub 2014 Jan 21.
Transplantation of bone marrow cells into spinal cord lesions promotes functional recovery in animal models, and recent clinical trials suggest possible recovery also in humans. The mechanisms responsible for these improvements are still unclear.
To characterize spinal cord motor neurite interactions with human bone marrow stromal cells (MSCs) in an in vitro model of spinal cord injury (SCI).
STUDY DESIGN/SETTING: Previously, we have reported that human MSCs promote the growth of extending sensory neurites from dorsal root ganglia (DRG), in the presence of some of the molecules present in the glial scar, which are attributed with inhibiting axonal regeneration after SCI. We have adapted and optimized this system replacing the DRG with a spinal cord culture to produce a central nervous system (CNS) model, which is more relevant to the SCI situation.
We have developed and characterized a novel spinal cord culture system. Human MSCs were cocultured with spinal motor neurites in substrate choice assays containing glial scar-associated inhibitors of nerve growth. In separate experiments, MSC-conditioned media were analyzed and added to spinal motor neurites in substrate choice assays.
As has been reported previously with DRG, substrate-bound neurocan and Nogo-A repelled spinal neuronal adhesion and neurite outgrowth, but these inhibitory effects were abrogated in MSC/spinal cord cocultures. However, unlike DRG, spinal neuronal bodies and neurites showed no inhibition to substrates of myelin-associated glycoprotein. In addition, the MSC secretome contained numerous neurotrophic factors that stimulated spinal neurite outgrowth, but these were not sufficient stimuli to promote spinal neurite extension over inhibitory concentrations of neurocan or Nogo-A.
These findings provide novel insight into how MSC transplantation may promote regeneration and functional recovery in animal models of SCI and in the clinic, especially in the chronic situation in which glial scars (and associated neural inhibitors) are well established. In addition, we have confirmed that this CNS model predominantly comprises motor neurons via immunocytochemical characterization. We hope that this model may be used in future research to test various other potential interventions for spinal injury or disease states.
在动物模型中，将骨髓细胞移植到脊髓损伤处可促进功能恢复，近期的临床试验表明人类也可能出现恢复情况。导致这些改善的机制仍不清楚。
在脊髓损伤（SCI）的体外模型中，描述脊髓运动神经突与人类骨髓基质细胞（MSC）之间的相互作用。
研究设计/背景：此前，我们报道过在存在胶质瘢痕中一些分子的情况下，人类MSC可促进背根神经节（DRG）中延伸的感觉神经突的生长，这些分子被认为在SCI后抑制轴突再生。我们对该系统进行了调整和优化，用脊髓培养物替代DRG以构建一个与SCI情况更相关的中枢神经系统（CNS）模型。
我们开发并描述了一种新型的脊髓培养系统。在含有与胶质瘢痕相关的神经生长抑制剂的底物选择试验中，将人类MSC与脊髓运动神经突共培养。在单独的实验中，分析MSC条件培养基，并将其添加到底物选择试验中的脊髓运动神经突中。
如先前关于DRG的报道，与底物结合的神经黏蛋白和Nogo - A排斥脊髓神经元的黏附以及神经突的生长，但在MSC/脊髓共培养中这些抑制作用被消除。然而，与DRG不同，脊髓神经元胞体和神经突对髓鞘相关糖蛋白的底物没有表现出抑制作用。此外，MSC分泌组包含许多刺激脊髓神经突生长的神经营养因子，但这些不足以在神经黏蛋白或Nogo - A的抑制浓度下促进脊髓神经突的延伸。
这些发现为MSC移植如何在SCI动物模型和临床中促进再生和功能恢复提供了新的见解，特别是在胶质瘢痕（及相关神经抑制剂）已充分形成的慢性情况下。此外，我们通过免疫细胞化学鉴定证实该CNS模型主要由运动神经元组成。我们希望该模型可用于未来研究，以测试针对脊髓损伤或疾病状态的各种其他潜在干预措施。
