論文

査読有り
2017年6月

Optimisation of murine organotypic slice culture preparation for a novel sagittal-frontal co-culture system

JOURNAL OF NEUROSCIENCE METHODS
  • Sarah Joost
  • ,
  • Kazuto Kobayashi
  • ,
  • Andreas Wree
  • ,
  • Stefan Jean-Pierre Haas

285
開始ページ
49
終了ページ
57
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1016/j.jneumeth.2017.05.003
出版者・発行元
ELSEVIER SCIENCE BV

Background: The nigrostriatal pathway is of great importance for the execution of movements, especially in the context of Parkinson's disease. In research, analysis of this pathway often requires the application of severe animal experiments. Organotypic nigrostriatal slice cultures offer a resource-saving alternative to animal experiments for research on the nigrostriatal system.
New method: We have established a time-saving protocol for the preparation of murine sagittal nigrostriatal slice cultures by using a tissue chopper and agarose embedding instead of a vibratome. Furthermore, we developed the first murine co-culture model and the first co-culture utilising sagittal slices for modelling the nigrostriatal pathway.
Results: Sagittal nigrostriatal slice cultures show good overall tissue preservation and a high number of morphologically unimpaired dopaminergic neurons in the substantia nigra. Sagittal-frontal co-culture demonstrates massive outgrowth of dopaminergic fibres from the substantia nigra into co-cultured tissue.
Comparison with existing methods: The use of a tissue chopper instead of a vibratome allows notable time-saving during culture preparation, therefore allowing optimisation of the preparation time. Sagittal co-cultures offer the opportunity to study dopaminergic fibres in their physiological environment and in co-cultured tissue from a different animal in the same culture system.
Conclusion: We here present a possibility to optimise the slice culture preparation process with the simple means of using a tissue chopper and fast agarose embedding. Furthermore, our sagittal-frontal co-culture system is suitable for the observation of dopaminergic outgrowth in both co-cultured tissues. (C) 2017 Elsevier B.V. All rights reserved.

リンク情報
DOI
https://doi.org/10.1016/j.jneumeth.2017.05.003
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/28478074
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000403995400006&DestApp=WOS_CPL
ID情報
  • DOI : 10.1016/j.jneumeth.2017.05.003
  • ISSN : 0165-0270
  • eISSN : 1872-678X
  • PubMed ID : 28478074
  • Web of Science ID : WOS:000403995400006

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