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将雪旺细胞和/或嗅鞘胶质细胞移植到挫伤的脊髓中:存活、迁移、轴突关联及功能恢复。

Transplantation of Schwann cells and/or olfactory ensheathing glia into the contused spinal cord: Survival, migration, axon association, and functional recovery.

作者信息

Pearse Damien D, Sanchez Andre R, Pereira Francisco C, Andrade Christian M, Puzis Raisa, Pressman Yelena, Golden Kevin, Kitay Brandon M, Blits Bas, Wood Patrick M, Bunge Mary Bartlett

机构信息

Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, USA.

出版信息

Glia. 2007 Jul;55(9):976-1000. doi: 10.1002/glia.20490.

Abstract

Schwann cells (SCs) and olfactory ensheathing glia (OEG) have shown promise for spinal cord injury repair. We sought their in vivo identification following transplantation into the contused adult rat spinal cord at 1 week post-injury by: (i) DNA in situ hybridization (ISH) with a Y-chromosome specific probe to identify male transplants in female rats and (ii) lentiviral vector-mediated expression of EGFP. Survival, migration, and axon-glia association were quantified from 3 days to 9 weeks post-transplantation. At 3 weeks after transplantation into the lesion, a 60-90% loss of grafted cells was observed. OEG-only grafts survived very poorly within the lesion (<5%); injection outside the lesion led to a 60% survival rate, implying that the injury milieu was hostile to transplanted cells and or prevented their proliferation. At later times post-grafting, p75(+)/EGFP(-) cells in the lesion outnumbered EGFP(+) cells in all paradigms, evidence of significant host SC infiltration. SCs and OEG injected into the injury failed to migrate from the lesion. Injection of OEG outside of the injury resulted in their migration into the SC-injected injury site, not via normal-appearing host tissue but along the pia or via the central canal. In all paradigms, host axons were seen in association with or ensheathed by transplanted glia. Numerous myelinated axons were found within regions of grafted SCs but not OEG. The current study details the temporal survival, migration, axon association of SCs and OEG, and functional recovery after grafting into the contused spinal cord, research previously complicated due to a lack of quality, long-term markers for cell tracking in vivo.

摘要

施万细胞(SCs)和嗅鞘胶质细胞(OEG)已显示出在脊髓损伤修复方面的潜力。我们试图在损伤后1周将其移植到成年大鼠挫伤的脊髓中后,通过以下方法在体内对它们进行识别:(i)使用Y染色体特异性探针进行DNA原位杂交(ISH),以识别雌性大鼠中的雄性移植细胞;(ii)慢病毒载体介导的EGFP表达。从移植后3天到9周对细胞存活、迁移以及轴突与胶质细胞的关联进行了定量分析。移植到损伤部位3周后,观察到移植细胞损失了60%-90%。仅OEG移植在损伤部位内存活情况非常差(<5%);在损伤部位外注射导致60%的存活率,这意味着损伤环境对移植细胞不利或阻止了它们的增殖。在移植后的后期,损伤部位的p75(+)/EGFP(-)细胞在所有实验范式中都多于EGFP(+)细胞,这是宿主施万细胞大量浸润的证据。注入损伤部位的SCs和OEG未能从损伤部位迁移。在损伤部位外注射OEG导致它们迁移到注射SCs的损伤部位,不是通过外观正常的宿主组织,而是沿着软脑膜或通过中央管。在所有实验范式中,都观察到宿主轴突与移植的胶质细胞相关联或被其包裹。在移植SCs的区域内发现了许多有髓轴突,但在OEG移植区域内未发现。本研究详细描述了SCs和OEG移植到挫伤脊髓后的时间存活、迁移、轴突关联以及功能恢复情况,此前由于缺乏用于体内细胞追踪的高质量长期标记物,该研究一直很复杂。

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