Spontaneous regeneration of intrinsic spinal cord axons in a novel spinal cord slice culture model
Spontaneous regeneration of intrinsic spinal cord axons in a novel spinal cord slice culture model
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DOI：
10.1111/j.1460-9568.2008.06227.x
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发表时间：
2008-05-01
影响因子：
3.4
通讯作者：
Kapfhammer, Josef P.
中科院分区：
文献类型：
--
作者：
Bonnici, Brenda;Kapfhammer, Josef P.
轴突修复研究中的一个重要问题是使用各种各样的损伤模型和与各自体内损伤范例相关的复杂性。器官型切片培养是一种潜在的体外替代方法，因为细胞结构组织组织保存良好，切片可以在培养中维持数周。迄今为止，还没有脊髓切片培养模型可用于轴突生长的研究。在这里，我们提出了一个脊髓切片培养模型，是非常适合轴突生长的研究。脊髓切片不是在横向而是在矢状纵向平面中切割，使得几个脊髓节段包括在切片培养物中。在这些文化中，典型的腹背极性的脊髓和内在的脊髓轴突形成了一个强大的纤维束延伸沿着切片的纵轴。在培养期间，轴突变成有髓鞘的，并且在这些培养物中存在突触接触。机械损伤后，脊髓内轴突具有轴突生长和再生的巨大潜力。随着培养物成熟度的增加，穿过病变部位的再生轴突的数量减少，但即使在成熟的培养物中，也存在少量的交叉纤维。这种切片培养模型可以为脊髓轴突生长和再生、突触形成、脊髓固有回路形成和髓鞘形成等方面的研究提供重要工具。
A substantial problem in research concerned with axonal repair is the use of a wide variety of lesion models and the complexity associated with the respective in vivo lesion paradigms. Organotypic slice cultures are a potential in vitro alternative because the cytoarchitectonic tissue organization is well preserved and the slices can be maintained in culture for several weeks. Until now no spinal cord slice culture model for the study of axonal growth has been available. Here we present a spinal cord slice culture model that is well suited for the study of axonal growth. The spinal cord slices were cut not in the transverse but in the sagittal longitudinal plane such that several spinal cord segments were included in the slice culture. In these cultures the typical ventro-dorsal polarity of the spinal cord was maintained and intrinsic spinal cord axons formed a strong fibre tract extending along the longitudinal axis of the slice. The axons became myelinated during the culture period and synaptic contacts were present in these cultures. After mechanical lesions the intrinsic spinal cord axons had a substantial potential for axonal growth and regeneration. The number of regenerating axons crossing the lesion site decreased with increasing maturation of the culture, but even in mature cultures a small number of crossing fibres were present. This slice culture model could provide an important tool for several aspects of spinal cord research in the fields of axonal growth and regeneration, synapse formation, formation of intrinsic spinal cord circuits and myelination.