Cord Blood is Neuroprotective in a Rat Model of Stroke

在美国，脑血管疾病是导致发病率和死亡率的第三大原因。作为 随着人口老龄化，开发有效的治疗方案将变得更加紧迫，不仅可以 提高存活率，但减少残疾。在卒中大鼠模型(大脑中动脉闭塞或 MCAO)我们发现，静脉注射(Iv)人脐带单个核组分 血液(HUCB)增强运动功能。在这项提案中，我们将探索如何管理这一 细胞群可能通过直接作用诱导MCAO的修复和减轻解剖损伤 神经保护和抗炎机制。在Aim 1WE中，我们将描述iv HUCB细胞如何 在给药24小时至7天后改变大脑中动脉阻塞的基本病理和炎症 MCAO使用炎症、神经元死亡、神经存活和细胞凋亡的标记物。在目标2中，我们将 确定特定的HUCB细胞亚群，它有助于诱导行为和 解剖学恢复。这将通过使用荧光激活细胞分选(FACS)来实现 用于移植到中风大鼠体内的细胞、B细胞、单核细胞和“干细胞”部分。中的端点度量 这些研究将包括一系列运动测试和脑梗塞体积的表现。目标3提出了一个 检测HUCB细胞是否改变大脑中动脉阻塞的炎症反应的一系列体外研究 通过与神经细胞群体的直接相互作用。用神经细胞，星形细胞， 少突胶质细胞或小胶质细胞将被用来识别直接相互作用和分子信号 介导缺氧/复氧反应中的存活、细胞因子表达和核因子-kappaB结合活性。 在目标4中，我们将使用微阵列技术来识别神经修复/存活基因，这些基因由 HUCB对神经元培养的治疗。所有的微阵列实验将通过蛋白质印迹和 免疫组织化学。这里提出的研究将增加我们对神经保护的理解 HUCB细胞诱导卒中康复的抗炎机制 并可能确定中风的其他潜在靶点，以开发治疗方案。 这项研究计划的最终目标是将一种创新的基于HUCB细胞的疗法开发成 移植治疗神经退行性疾病和脑损伤的可行临床选择。

Cerebrovascular disease is the third leading cause of morbidity and mortality in the United States. As the population ages, it will be even more pressing to develop effective treatment options that can not only increase survival, but decrease disability. In a rat model of stroke (the middle cerebral artery occlusion or MCAO) we found that intravenously (iv) administering the mononuclear fraction from human umbilical cord blood (HUCB) enhances motor functions. In this proposal we will explore how the administration of this population of cells may induce recovery and decrease anatomical damage induced by MCAO through direct neuroprotective and anti-inflammatory mechanisms. In Aim 1we will characterize how iv HUCB cells modify the underlying pathology and inflammation of the MCAO when administered 24 hours to 7 days after the MCAO using markers of inflammation, neuronal death, neurosurvival, and apoptosis. In Aim 2, we will identify the specific subpopulation of the HUCB cells that is instrumental in inducing behavioral and anatomical recovery. This will be accomplished using fluorescent activated cell sorting (FACS) to enrich T cell, B Cell, monocyte and "stem" cell fractions for transplantation into a stroked rat. Endpoint measures in these studies will include performance on a battery of motor tests and infarct volume. Aim 3 proposes a series of in vitro studies to examine whether the HUCB cells modify the inflammatory response to the MCAO through direct interactions with neural cell popultions. Cell culture assays using neuronal, astrocytic, oligodendrocytes or microglia will be conducted to identify direct interactions and the molecular signals that mediate survival, cytokine expression and NF-kappaB binding activity in response to hypoxia/reoxygenation. In Aim 4 we will use microarray technology to identify neural repair/survival genes that are up-regulated by HUCB treatment of neuronal cultures. All microarray experiments will be verified with western blots and immunohistochemistry. The studies proposed here will increase our understanding of the neuroprotective and anti-inflammatory mechanisms underlying the recovery from stroke induced by HUCB cell transplantation and may also identify other potential targets in stroke for development of treatment options. The ultimate goal of this research program is to develop an innovative HUCB cell based therapy into a viable clinical option for transplantation in neurodegenerative disease and brain injury.