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

Shortiss, Ciara, Howard, Linda, & McMahon, Siobhan S. (2025). Viability and suitability of rat pup organotypic spinal cord slice cultures in a transection injury model. Journal of Neuroscience Methods, 421, 110499. https://doi.org/10.1016/j.jneumeth.2025.110499

Abstract

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.

Funder

Irish Research Council
College of Medicine, Nursing and Health Science, University of Galway

Publisher

Elsevier

Rights

Attribution 4.0 International

cb

Collections

Anatomy (Scholarly Articles)