首页> 外文期刊>Journal of Neuroscience Methods >Genetically modified canine Schwann cells--In vitro and in vivo evaluation of their suitability for peripheral nerve tissue engineering.
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Genetically modified canine Schwann cells--In vitro and in vivo evaluation of their suitability for peripheral nerve tissue engineering.

机译:转基因犬雪旺细胞-体外和体内评估其对周围神经组织工程的适用性。

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摘要

After peripheral nerve injury, Schwann cells (SC) guarantee for a regeneration-promoting milieu and are crucially involved in axonal regeneration. For extended nerve defects, bridging with an autologous nerve transplant is the gold standard therapy. Artificial biohybrid nerve transplants which combine a synthetic conduit with autologous SC genetically modified to express regeneration-promoting proteins may provide an alternative therapy to autotransplantation. The dog seems to be an ideal translational animal model for new treatment strategies. In the present study, utilizing a new transfection protocol, we transplanted enhanced green fluorescent protein (EGFP)-expressing adult canine SC (cSC) into a 5mm epineural pouch in the sciatic nerve of adult rats (n=9). The epineurial pouch technique serves as proof of principle to follow the fate of the transplanted cSC for up to 14 days after surgery. Fluorescence microscopy and immunohistochemistry revealed survival and integration of EGFP-expressing cSC into the regenerating host nerve tissue. We demonstrate that transplanted cSC contribute to the formation of bands of Bungner and are located in close vicinity to growth-associated protein-43 (GAP-43) expressing regenerating nerve fibers. This provides first evidence that transplanted genetically modified Schwann cells do successfully integrate into the host tissue where they could actively contribute to the regeneration process.
机译:周围神经损伤后,许旺细胞(SC)保证了促进再生的环境,并参与了轴突的再生。对于延伸的神经缺损,采用自体神经移植架桥是金标准疗法。人工生物杂交神经移植将合成导管与经过基因修饰的自体SC结合在一起,以表达促进再生的蛋白质,可以为自体移植提供替代疗法。狗似乎是新治疗策略的理想转化动物模型。在本研究中,我们利用新的转染方案,将表达增强型绿色荧光蛋白(EGFP)的成年犬SC(cSC)移植到成年大鼠坐骨神经(n = 9)的5mm硬膜外囊中。手术后长达14天,肾小管囊技术可以作为原则证明移植的cSC的命运。荧光显微镜和免疫组织化学揭示了表达EGFP的cSC的存活和整合到再生宿主神经组织中。我们证明,移植的cSC有助于形成的Bungner乐队,并位于与生长相关的蛋白43(GAP-43)表达再生神经纤维的紧邻。这提供了第一个证据,证明移植的经过基因修饰的雪旺氏细胞确实成功整合到宿主组织中,在那里它们可以积极地促进再生过程。

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