Mechanical Distention of Organotypic Hippocampal Slice Culture Membranes for High-Throughput In Vitro Modeling of Traumatic Brain Injury

Authors

• Julia Jagielo-Miller, University of KentuckyFollow
• Caleb Bailey, University of Kentucky College of MedicineFollow
• Samir Patel, University of KentuckyFollow
• Paresh Prajapati, University of Kentucky College of Medicine
• Cassidy Leibold, University of KentuckyFollow
• Patrick Sullivan, University of KentuckyFollow
• Mark Prendergast, University of KentuckyFollow

Abstract

Traumatic brain injury (TBI) results from impact to the head that induces both primary and secondary injuries. Secondary injuries are characterized by downstream inflammation, metabolic dysfunction, and cell death manifest from the inflicting primary injury to the head. Secondary injury offers a window for therapeutic interventions, but the multifaceted nature of secondary injury is complicated, necessitating mechanistic tools to screen the efficacy of such interventions. As such, utilizing animal models to define the features of secondary injury mechanisms is critical for medications development. Various animal TBI models employ specialized equipment to recapitulate both primary and secondary injury aspects of human TBI. The organotypic hippocampal slice culture (OHSC) model offers a biological intermediate between live animal and dissociated cell culture models. In OHSC models, ex vivo tissue containing heterogenous hippocampal cell types is plated upon permeable culture membranes, which have the capacity to be manipulated. We, therefore, repurposed a commercially available impact device to mechanically distend the OHSC culture membrane, effectively inducing an indirect stretch injury to hippocampal tissue. This stretch injury technique causes characteristic secondary injury trauma, such as widespread cell death, loss of neuronal viability, and production of reactive oxygen species, following the initial insult. Importantly, both the impact force and dwell time of the membrane distention are scalable, a modular feature widely employed across other animal TBI models. This OHSC TBI model may lend itself to high-throughput preliminary assessment of therapeutic efficacy for treatment of secondary injury in animal TBI models.

Document Type

Article

Publication Date

10-1-2025

Notes/Citation Information

Publisher Copyright: © 2025 The Author(s)

Digital Object Identifier (DOI)

10.1177/2689288X251389788

Archival?

Archival

Repository Citation

Jagielo-Miller, Julia; Bailey, Caleb; Patel, Samir; Prajapati, Paresh; Leibold, Cassidy; Sullivan, Patrick; and Prendergast, Mark, "Mechanical Distention of Organotypic Hippocampal Slice Culture Membranes for High-Throughput In Vitro Modeling of Traumatic Brain Injury" (2025). Biology Faculty Publications. 250.
https://uknowledge.uky.edu/biology_facpub/250