Stroke therapy using embryonic stem cells and bone marrow stromal cell-derived neuralstem cells

骨髓基质细胞(MSCs)是治疗多种中枢神经系统疾病的良好细胞来源。我们报道了通过导入含有Notch-1细胞内域的细胞，然后在自由培养系统中生长，有效地从大鼠和人类MSCs(NS-MSCs)诱导出定向神经前体细胞。为了确定NS-MSCs的治疗潜力，在大鼠局灶性脑缺血模型中，将细胞移植到皮质和纹状体。移植的NS-MSCs在宿主脑内的存活率、分布和整合率都很高，在第100天，移植的NS-MSCs的多巴胺能、谷氨酸能和γ-氨基丁酸(GABA)能神经元标记呈阳性。他们将长的神经突起延长近6.3毫米，其中许多表达突触素。在肢体放置和水迷宫试验中也观察到显著的行为恢复。建立具有自我…的胚胎干细胞治疗脑缺血的细胞疗法为了获得更多的尤文和多能性，我们采用无血清悬浮培养的方法诱导小鼠ES细胞分化为神经前体细胞，并在体外和体内证实了各种基础神经节神经元标志物和神经递质相关标志物的表达，被认为适合于修复大脑中动脉闭塞后受损的纹状体。我们通过荧光激活细胞分选纯化了表达神经前体标志物Sox1的神经前体细胞，并对Sox1阳性神经前体细胞进行了2个月的预防缺血脑内肿瘤形成的实验。我们还对移植细胞的存活和分化以及缺血损伤后的功能恢复进行了分析。诱导的多能干细胞具有类似于胚胎干细胞的自我更新和多潜能的特性。它们是细胞替代疗法合适的细胞来源的一个很好的候选者。在这项研究中，我们将人iPS细胞衍生的神经前体细胞移植到缺血小鼠的大脑中。采用类胚体集合体(SFEB)无血清培养方法，将人iPS细胞分化为神经前体细胞。缺血诱导后1周将供体细胞移植到缺血侧纹状体内。细胞在移植部位存活，部分细胞沿外囊和胼胝体迁移。移植组大鼠的行为恢复明显增强。我们的结果提示，人iPS细胞来源的神经前体细胞在缺血脑内存活和迁移，并有助于神经回路重建的功能恢复。安全有效地将胚胎干细胞移植到脑内需要抑制对同种异体移植的局部炎症和免疫反应。为了研究影响移植细胞存活和分化的细胞因子，我们利用基质细胞来源的诱导活性来诱导小鼠ES细胞分化为神经前体细胞，并将其移植到小鼠脑内。对周围脑组织的检查显示，鼻咽癌移植后IL-1β、IL-4和IL-6的表达水平升高。其中，只有IL-6在体外抑制神经元分化，促进神经胶质分化。当我们在移植过程中将抗IL-6受体抗体添加到NC中时，这种单一的局部阻断IL-6信号的作用减少了宿主来源的白细胞的积累，包括小胶质细胞。此外，它还促进神经干细胞向神经元的分化，减少神经胶质细胞的分化，其程度与环孢素A全身持续给药相似。这些结果表明，在细胞替代疗法治疗神经系统疾病的过程中，抗IL-6受体抗体与神经干细胞联合应用可能会促进神经元的分化。较少

Bone marrow stromal cells(MSCs) are an excellent source of cells for treating a variety of central nervous system diseases. We report the efficient induction of committed neural progenitor cells from rat and human MSCs(NS-MSCs) by introduction of cells with the intracellular domain of Notch-1 followed by growth in the free-floating culture system. To determine the therapeutic potential of NS-MSCs, cells were transplanted into the cortex and striatum in a rat model of focal cerebral ischemia. The survival, distribution, and integration of NS-MSCs in the host brain were very high, and at day 100, grafted NS-MSCs were positive for dopaminergic, glutamatergic, and gamma-amino butyric acid(GABA) ergic neuronal markers. They extended long neurites for nearly 6. 3 mm and many of these expressed synaptophysin. Significant behavioral recovery was also observed in limb-placing and water-maze tests.To establish cell therapy for cerebral ischemia using embryonic stem(ES) cells, which have self-ren … More ewing and pluripotent capacities, we induced the differentiation of the neural progenitors from mouse ES cells using the serum-free suspension culture method and confirmed the expression of various basal ganglial neuronal markers and neurotransmitter-related markers both in vitro and in vivo, which was thought to be suitable for replacing damaged striatum after middle cerebral artery occlusion. We purified the progenitors expressing the neural progenitor marker Sox1 by fluorescence-activated cell sorting and Sox1-positive neural progenitors prevented tumor formation in ischemic brain for 2 months. We also analyzed survival and differentiation of transplanted cells and functional recovery from ischemic damage.Induced pluripotent stem(iPS) cells possess the properties of self-renewal and pluripotency, similar to embryonic stem cells. They are a good candidate as a source of suitable cells for cell replacement therapy. In this study, we transplanted human iPS cell-derived neural progenitors into an ischemic mouse brain. Human iPS cells were differentiated into neuronal progenitors by serum-free culture of embryoid body-like aggregates(SFEBs). Donor cells were transplanted into the ischemic lateral striatum 1week after ischemia induction. Cells survived at the transplantation site, with migration of a proportion of cells along the external capsule and corpus callosum. Behavioral recovery was significantly enhanced in the transplanted group. Our results suggest that human iPS cell-derived neuronal progenitors survive and migrate in the ischemic brain, and contribute toward functional recovery vianeural circuit reconstitution.Safe and efficient transplantation of embryonic stem(ES) cells to the brain requires that local inflammatory and immune responses to allogeneic grafts are inhibited. To investigate cytokines that affect graft cell survival and differentiation, we used stromal cell-derived inducing activity to induce the differentiation of neural progenitor cells(NPCs) from mouse ES cells and transplanted the NPCs into mouse brain. Examination of surrounding brain tissue revealed elevated expression levels of interleukin(IL)-1β, IL-4, and IL-6 in response to NPC transplantation. Among these, only IL-6 reduced neuronal differentiation and promoted glial differentiation in vitro. When we added anti-IL-6 receptor antibodies to NPCs during transplantation, this single and local blockade of IL-6 signaling reduced the accumulation of host-derived leukocytes, including microglia. Furthermore, it also promoted neuronal differentiation and reduced glial differentiation from the grafted NPCs to an extent similar to that with systemic and continuous administration of cyclosporine A. These results suggest that local administration of anti-IL-6 receptor antibodies with NPCs may promote neuronal differentiation during the treatment of neurological diseases with cell replacement therapy. Less