Long-Term Culture of Organotypic Hippocampal Slice from Old 3xTg-AD Mouse: An ex vivo Model of Alzheimer’s Disease

Abstract

Objective Conventional methods for organotypic hippocampal tissue slice culture (OHSC) have shown several disadvantages or limitations regarding age of animals used, duration of culture and difficulty using neurodegenerative models. Therefore, we tried to establish OHSC from old 3xTg-Alzheimer’s disease (AD) mice for longer period (over 4 weeks) and to validate utility of this system as a valid platform for translational neuroscience of AD. Methods OHSC was performed with old 3xTg-AD mice (12–14 months), old wild type mice (12–14 months) and young 3xTg-AD mice (2–4 months) using serum-free medium for 4 weeks. Hippocampal structure was evaluated by 4’, 6-diamidino-2-phenylindole (DAPI) intensity and neuronal metabolism was measured by Alamarblue assay. Pathologic characteristics of AD were also investigated; β-amyloid levels by ELISA, amyloid plaque deposition by Thioflavin-S staining, and glial activation by immunohistochemistry. Results Following 4-week culture in serum-free media, hippocampal cells and layers were well preserved in cultured slices from old AD mice as was in those from young AD and old wild type mice. On the contrary, excessive regression of total visible cells was observed in conventional serum-containing medium regardless of genotype of mice. In parallel with this well preserved structure, major pathologic characteristics of AD were also well manifested in hippocampal slices from old AD mice. Conclusion Our findings suggest that long-term OHSC from old 3xTg-AD mouse can serve as a promising ex vivo system for studies on pathophysiology of AD, especially with the minimum number of sacrifice of experimental animals.

Neuroprotective Effects of Kinin B2 Receptor in Organotypic Hippocampal Cultures of Middle-Aged Mice

“…Recently, Jang et al (2018) established organotypic cultures derived from old transgenic animals to study AD, where the slices are grown in serum-free media for 4 weeks. In our work, we established OHCs from 6- and 12-months-old C57Bl/6 animals with serum and demonstrated the ability of bradykinin to maintain viable cultures over a long period.…”

Fructose-1,6-Bisphosphate Protects Hippocampal Rat Slices from NMDA Excitotoxicity

“…The phenomenon of glutamate-mediated excitotoxicity plays a central role in acute and chronic neurodegenerations and represents a target for new therapeutic approaches in numerous pathological states, including cerebral ischemia, stroke, traumatic brain injury [35,36,37]. Organotypic hippocampal slice cultures represent a useful in vitro model with which to study not only the effects of excitotoxicity (triggered by the challenge with NMDA) on nervous cell metabolism and functions [38] but also to evaluate efficacy of drug treatments [39].…”

POSCAbilities: The Application of the Prion Organotypic Slice Culture Assay to Neurodegenerative Disease Research

“…Specifically, Humpel suggests culturing thinner slices (100–150 µm) from adults [ 82 ]. Other authors have found that adult mouse brain slices have greater viability with serum-free medium [ 83 , 84 ]. Treatment of slices with brain-derived neurotrophic factor has also proven beneficial [ 85 ].…”