Exper imenta I study of reconstruct ion of peripheral nerve defect using tissue-engineered bioabsorbable nerve conduit with iPS cell induced neurosphere cell

iPS细胞诱导神经球细胞组织工程生物可吸收神经导管重建周围神经缺损的实验研究

• 批准号: 21591904
• 负责人: TAKAMATSU Kiyohito
• 金额: $ 2.83万
• 依托单位: Osaka City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21591904/
• 关键词: 再生医療; 末梢神経; 再生; 生体吸収性材料; iPS細胞; 人工神経; 組織工学; マウス; 生体材料

【Background】in spite ofextensirosearcliof induced pluripotent stem(iPS) cells, the therapeutic potential of iPS cells for peripheral nerve injury is largely unknown. This is the study to prove the feasibility of the combination of iPS cell-derived neurospheres and bioabsorbable nerve conduit.【Purpose】<Aiml> The first purpose of this study was to examine the adhesion of the iPS cell induced neurospheres to the bioabsorbable nerve conduits and to examine their cellular characteristics.<Aim2> The second purpose of this study wasto test tissue-engineered hioabsorbable nerve conduits coated with a three-dimensional(3D) culture of WS cell-derived neurospheres in peripheral nerve repair in vivo.【Methods】<Aiml> The nerve conduit has 2 layers, The outer layer is composed of PLLA mesh and the inner layer is composedofPLLAand PCL porous sponge. We generated primary and secondary neurospheres from iPS cells by a published protocol[Editor1]. The primary and secondary neurospheres were suspended in … More each conduit. After suspension, the conduits were placed in a cell incubator for 7 or 14 days. After the conduits were seeded with iPs cells, they were subjected to immunohistological analysis by using antibodies specific to the glial marker(GFAP), Schwann cell marker(S-100) and neuronal marker.<Aim2> The secondary neurospheres derived from mouse iPS cells were suspended in each conduit.(4.0×10^6 cells per miduit) and cultured in 3D-cultu fir 14 days. We then implanted them in the mouse sciatic nerve gap(5 mm)(iPs group ; n=10). The nerve conduit. alone was implanted in the control group(n=10). Motor and sensory function recovery was assessed. At 4, 8, 12 weeks, nerve regeneration in the nerve conduit was evaluated by histological analysis.【Results】<Aiml> All the primary and secondary neurospheres that had differentiated for 7 or 14 days were found to have adhered to the inner surface of the conduits and migrated into the inner porous sponge. All neurospheres were positive for S-100 and GFAP but were negative for neurofilament protein. The cell adhesion and the immunostaining characteristics between the 7-and 14-day-differentiated neurospheres were not different.<Aim2> Motor and sensory function recovery was significantly faster in the iPS group at weeks 4, 8, and 12.At 12 weeks, all the nerve conduits remained structurally stable without any collapse. Histological analysis indicated axonal regeneration in the nerve conduits of both groups. However, the iPS group showed more vigorous axonal regeneration.【Conclusion】The bioabsorbable nerve conduits created by 3D culture of iPS cell-derived neurospheres promoted regeneration of peripheral nerves and functional recovery in vivo. The combination of iPS cell technology and bioabsorbable nerve conduits could represent a future tool for the treatment of peripheral nerve defects. Less

[背景]尽管有炎症性诱导的多能茎（IPS）细胞，但IPS细胞对周围神经损伤的治疗潜力在很大程度上尚不清楚。这是证明IPS细胞衍生的神经球和生物吸附神经导管的可行性的研究。 [Purpose] <Aiml> The first purpose of this study was to examine the adhesion of the iPS cell induced neurospheres to the bioabsorbable nerve conduits and to examine their cellular Characteristics.<Aim2> The second purpose of this study was tested tissue-engineered hioabsorbable nerve conduits coated with a three-dimensional (3D) culture of WS cell-derived neurospheres in peripheral nervous在体内修复。[方法] <AIML>神经导管具有2层，外层由PL​​LA网格组成，并且内层组成了Pllaand PCL多孔赞助商。我们通过公开的方案[Editor1]从IPS细胞中产生了初级和次级神经球。将主要和次级神经球悬挂在……每个导管中。悬浮后，将导管放在细胞孵化器中7或14天。导管与IPS细胞接种后，通过使用特异性的胶质标记物（GFAP），Schwann细胞标记（S-100）和神经元标记物进行免疫组织学分析。<AIM2>二级神经球源是从小鼠IPS细胞中衍生的二级神经球均在每个Codlit中悬浮在小鼠IPS中。然后，我们将它们植入小鼠坐骨神经间隙（5 mm）（IPS组； n = 10）。神经管道。单独植入对照组（n = 10）。评估了运动和感觉函数恢复。在4、8、12周时，通过组织学分析评估了神经导管中的神经再生。结果发现，发现了7或14天的所有主要和次级神经球均被发现已粘附到导管的内表面并迁移到内部多孔的孔子中。所有神经球均为S-100和GFAP阳性，但神经丝蛋白为阴性。 7和14天分差的神经球之间的细胞粘附和免疫染色特性没有差异。<AIM2>运动和感官功能恢复在IPS组中在第4、8和12周的IPS组中的速度明显更快。在12周，第12周，所有神经管道在没有任何崩溃的情况下保持结构稳定。组织学分析表明两组神经管道中的轴突再生。但是，IPS组显示出更剧烈的轴突再生。 [结论]由IPS细胞衍生的神经球的3D培养产生的生物吸附神经管道促进了周围神经的再生和体内功能恢复。 IPS细胞技术和可生物吸附神经管道的结合可能代表了治疗周围神经缺陷的未来工具。较少的

项目成果

ips細胞を用いたハイブリッド型人工神経の作成～人工神経上でのiPS細胞培養

使用 ips 细胞创建混合型人工神经 ~ 在人工神经上培养 iPS 细胞

• 发表时间: 2010
• 作者: 松山大輔;渡辺雅彦;隅山香織;黒岩真弘;持田讓治;高松聖仁
• 通讯作者: 高松聖仁