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Psychiatry Investig > Volume 15(2); 2018 > Article
Psychiatry Investigation 2018;15(2):205-213.
DOI: https://doi.org/10.30773/pi.2017.04.02    Published online November 29, 2017.
Long-Term Culture of Organotypic Hippocampal Slice from Old 3xTg-AD Mouse: An ex vivo Model of Alzheimer’s Disease
Sooah Jang1,2, Hyunjeong Kim1,3, Hye-jin Kim1, Su Kyoung Lee1, Eun Woo Kim1,3, Kee Namkoong1,2, Eosu Kim1,2,3
1Department of Psychiatry, Yonsei University College of Medicine, Seoul, Republic of Korea
2Institute of Behavioral Science in Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea
3Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea
Correspondence  Eosu Kim ,Tel: +82-2-2228-1620, Fax: +82-2-313-0891, Email: kimeosu@yuhs.ac
Received: January 7, 2017   Revised: March 14, 2017   Accepted: April 2, 2017   Published online: November 29, 2017
*Sooah Jang and Hyunjeong Kim contributed equally to this study as co-first authors.
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.
Keywords: Organotypic slice culture, Ex vivo, Hippocampus, Alzheimer’s disease
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