大鼠幼鼠器官型脊髓切片培养在横断损伤模型中的可行性和适用性

IF 2.7 4区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Neuroscience Methods Pub Date : 2025-05-24 DOI:10.1016/j.jneumeth.2025.110499

Ciara Shortiss , Linda Howard , Siobhan S. McMahon

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引用次数: 0

摘要

活体器官型脊髓切片培养（OSC）模型有利于脊髓损伤（SCI）的研究。它们保留了体内细胞的相互作用，但涉及较少的伦理问题，并且比体内模型更容易操作。由于早期产后动物的生存能力提高，OSCs是首选。从出生第4天(P)大鼠获得的横断损伤OSC模型已经得到验证，然而，由于啮齿动物脊髓发育在出生后早期发生了显著变化，因此年龄较大的出生第4天大鼠可能更适合OSC模型。新方法本研究比较了P11大鼠OSCs与建立的P4横断模型的生存能力。结果各年龄组P4与P11对照切片、对照与损伤切片间细胞活力均无显著差异。与P4 osc相比，P11 osc在损伤后也表现出强大的活细胞长入横断损伤间隙，并增加了沿横断间隙边缘胶质疤痕标记物GFAP和CSPGs的免疫染色体积分数。没有发现性别对这些参数有任何影响。与现有方法比较，我们的研究结果表明，P11 osc在横断损伤后保持活力和细胞长入，与已建立的P4衍生osc相当。P11 osc也被发现在横断损伤后表现出胶质瘢痕反应。结论来自P11动物的soscs维持了与P4 OSCs相当的活力，同时也表现出疤痕反应。本研究认为P11 OSC是P4 OSC横断性SCI模型的有效替代。

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Viability and suitability of rat pup organotypic spinal cord slice cultures in a transection injury model

Background

Ex vivo organotypic spinal cord slice culture (OSC) models are advantageous for spinal cord injury (SCI) research. They retain in vivo cellular interactions but involve fewer ethical concerns and are easier to manipulate than in vivo models. Due to improved viability OSCs derived from early postnatal animals are preferred. A transection injury OSC model derived from postnatal day (P) 4 rats has been validated, however as rodent spinal cord development changes significantly in early postnatal weeks, older postnatal day animals may be more appropriate for OSC models.

New method

This study compared the viability of OSCs derived from P11 rats with the established P4 transection model.

Results

No significant differences in cell viability were found between P4 and P11 control slices, or between control and injured slices within each age group. P11 OSCs also demonstrated strong viable cell ingrowth into a transection injury gap post-injury, comparable to P4 OSCs, and increased their volume fraction of immunostaining for the glial scar markers GFAP and CSPGs along transection gap edges. Sex was not found to affect any of these parameters.

Comparison with existing methods

Our findings demonstrate that P11 OSCs maintain viability and cellular ingrowth post transection injury comparable to that of the well-established P4 derived OSCs. P11 OSCs were also found to exhibit a glial scarring response post transection injury.

Conclusions

OSCs from P11 animals maintain viability comparable to P4 OSCs while also exhibiting a scarring response. This study concludes that P11 OSCs are a valid alternate to P4 OSC transection SCI model.

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来源期刊

Journal of Neuroscience Methods 医学-神经科学

CiteScore

7.10

自引率

3.30%

发文量

226

审稿时长

52 days

期刊介绍： The Journal of Neuroscience Methods publishes papers that describe new methods that are specifically for neuroscience research conducted in invertebrates, vertebrates or in man. Major methodological improvements or important refinements of established neuroscience methods are also considered for publication. The Journal''s Scope includes all aspects of contemporary neuroscience research, including anatomical, behavioural, biochemical, cellular, computational, molecular, invasive and non-invasive imaging, optogenetic, and physiological research investigations.