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

使用胚胎干细胞和骨髓基质细胞衍生的神经干细胞治疗中风

• 批准号: 20390382
• 负责人: TAKAGI Yasushi
• 金额: $ 12.31万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20390382/
• 关键词: 脳梗塞; 神経幹細胞; 骨髄間質細胞; 胚性幹細胞; 移植; ES細胞; iPS細胞; 脳虚血; 移植治療; 脳血管障害学; 神経幹細胞移植

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

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。我们报告了通过引入具有Notch-1细胞内结构域的细胞，然后在自由漂浮培养系统中生长，从大鼠和人MSC（NS-MSC）有效诱导定向神经祖细胞。为了确定NS-MSCs的治疗潜力，将细胞移植到大鼠局灶性脑缺血模型的皮质和纹状体中。NS-MSC在宿主脑中的存活、分布和整合非常高，并且在第100天，移植的NS-MSC对多巴胺能、多巴胺能和γ-氨基丁酸（GABA）能神经元标记物呈阳性。他们延长长神经突近6。3 mm，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。利用具有自我修复能力的胚胎干细胞（ES细胞）建立脑缺血的细胞治疗方法， ...更多信息 本研究以小鼠胚胎干细胞为研究对象，利用无血清悬浮培养法诱导小鼠胚胎干细胞向神经前体细胞分化，并在体内外证实了多种基底节神经元标志物和神经递质相关标志物的表达，认为其适合于替代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表达神经祖细胞标记物Sox 1的祖细胞，Sox 1阳性神经祖细胞在缺血性脑中预防肿瘤形成2个月。诱导性多能干细胞（induced pluripotent stem，iPS）具有与胚胎干细胞相似的自我更新和多能性的特性。它们是细胞替代疗法的合适细胞来源的良好候选者。在这项研究中，我们将人iPS细胞衍生的神经祖细胞移植到缺血小鼠脑中。人iPS细胞通过胚状体样聚集体（SFEBs）的无血清培养分化为神经元祖细胞。缺血1周后将供体细胞移植到缺血侧纹状体内。细胞在移植部位存活，部分细胞沿着外囊和胼胝体迁移。移植组的行为恢复明显增强。我们的研究结果表明，人iPS细胞来源的神经元祖细胞在缺血脑中存活和迁移，并通过神经回路重建促进功能恢复。安全有效的胚胎干细胞（ES细胞）脑移植需要抑制对同种异体移植物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，我们使用基质细胞衍生的诱导活性来诱导小鼠ES细胞向神经祖细胞（NPC）分化，并将NPC移植到小鼠脑中。对周围脑组织的检查显示，NPC移植后白细胞介素（IL）-1β、IL-4和IL-6的表达水平升高。其中，只有IL-6在体外减少神经元分化和促进胶质细胞分化。当我们在移植过程中向NPC中加入抗IL-6受体抗体时，这种IL-6信号的单一和局部阻断减少了宿主来源的白细胞（包括小胶质细胞）的积累。此外，它还促进神经元分化和减少神经胶质细胞分化的移植的NPC的程度类似于与环孢素A的全身和连续给药。这些结果表明，在用细胞替代疗法治疗神经系统疾病期间，局部施用抗IL-6受体抗体与NPC可促进神经元分化。少

项目成果

胚性幹細胞、人工多能性幹細胞由来神経幹細胞の虚血脳への移植-現状と課題について-

将胚胎干细胞和诱导多能干细胞衍生的神经干细胞移植到缺血脑中 - 现状和挑战 -

• 发表时间: 2012
• 作者: Kikuchi Y;Yasuhara T;Agari T;Kondo A;Kuramoto S;Kameda M;Kadota T;Baba T;Tajiri N;Wang F;Tayra JT;Liang H;Miyoshi Y;Borlongan CV;Date I;高木康志
• 通讯作者: 高木康志

Tissue inhibitor of metalloproteinases protect blood-brain barrier disruption in focal cerebral ischemia

• DOI: 10.1038/jcbfm.2008.59
• 发表时间: 2008-10-01
• 期刊: JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM
• 影响因子: 6.3
• 作者: Fujimoto, Motoaki;Takagi, Yasushi;Nozaki, Kazuhiko
• 通讯作者: Nozaki, Kazuhiko

Functional recovery of the murine brain ischemia model using human induced pluripotent stem cell-derived telencephalic progenitors

• DOI: 10.1016/j.brainres.2012.03.049
• 发表时间: 2012-06-12
• 期刊: BRAIN RESEARCH
• 影响因子: 2.9
• 作者: Gomi, Masanori;Takagi, Yasushi;Takahashi, Jun
• 通讯作者: Takahashi, Jun

Generate integration-free human iPS cells

生成免整合的人类 iPS 细胞

• 发表时间: 2011
• 期刊: Nat Methods
• 影响因子: 48
• 作者: Okita K;Matsumura Y;Sato Y;Okada A;Morizane A;Okamoto S;Hong H;Nakagawa M;Tanabe K;Tezuka K;Shibata T;Kunisada T;Takahashi M;Takahashi J;Saji H;Yamanaka S.
• 通讯作者: Yamanaka S.

Committed neural progenitor cells derived from genetically modified bone marrow stromal cells ameliorate deficits in a rat model of stroke

• DOI: 10.1038/jcbfm.2009.62
• 发表时间: 2009-08-01
• 期刊: JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM
• 影响因子: 6.3
• 作者: Hayase, Makoto;Kitada, Masaaki;Dezawa, Mari
• 通讯作者: Dezawa, Mari