イシヅカ　ユウタ   Yuta Ishizuka   石塚　佑太    所属   川崎医科大学  医学部 基礎医学　病態代謝学    職種   講師
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	A simple DMSO-based method for cryopreservation of primary hippocampal and cortical neurons.
掲載誌名	正式名：Journal of neuroscience methods 略　 称：J Neurosci Methods ISSNコード：1872678X/01650270
掲載区分	国外
出版社	ELSEVIER
巻・号・頁	333,pp.108578
著者・共著者	Yuta Ishizuka*, Clive R. Bramham*
担当区分	筆頭著者,責任著者
発行年月	2020/03
概要	BACKGROUND:Primary neuronal cultures are widely used to elucidate fundamental aspects of neuronal anatomy, physiology, cell biology, and neuronal dysfunction in animal models of disease. However, preparation of primary neuronal cultures from rodent embryos is labor-intensive, and it is often difficult to produce high-quality cultures consistently in a single laboratory, and to compare results between laboratories. To overcome these issues, cryopreservation can be used to obtain more standardized, high-quality banks of neuronal cultures.NEW METHOD:In this study, we present a simplified cryopreservation method for rodent primary hippocampal and cortical neurons from embryonic day 18.5 fetuses, using DMSO-containing traditional cell freezing medium.RESULTS:Cryopreserved neurons stored for more than 1 year in liquid nitrogen were assessed by cell imaging, as well as biochemical signaling transduction and gene expression in response to pharmacological treatments. Cryopreserved neuronal cultures were comparable to freshly prepared cultures in terms of: (1) neuronal viability, (2) neuronal morphology and maturation, (3) functional synapse formation, (4) stimulus responsiveness. These results indicate that DMSO-cryopreserved neurons are equivalent to freshly prepared neurons both developmentally and functionally.COMPARISON WITH EXISTING METHODS:Our method is simple and does not require special reagents or equipment.CONCLUSIONS:Introduction of the cryopreserved neurons as a standard laboratory practice has the potential to increase the robustness and reproducibility of findings between laboratories and reduce the number of animals used in research.
DOI	10.1016/j.jneumeth.2019.108578
PMID	31899209