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Dissection of mouse hippocampus and preparation of single cell suspension

AC Arianna Casciati
KD Katalin Dobos
FA Francesca Antonelli
AB Anett Benedek
SK Stefan J. Kempf
MB Montserrat Bellés
AB Andrea Balogh
MT Mirella Tanori
LH Luis Heredia
MA Michael J. Atkinson
CT Christine von Toerne
OA Omid Azimzadeh
AS Anna Saran
GS Geza Sáfrány
MB Mohammed A. Benotmane
ML M. Victoria Linares-Vidal
ST Soile Tapio
KL Katalin Lumniczky
SP Simonetta Pazzaglia
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After cervical dislocation the hippocampus was dissected from whole brains. Hippocampal tissue was finely chopped, followed by enzymatic digestion, which was done with the Miltenyi Neural Dissociation Kit (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) according to the manufacturer's instructions. To separate the cellular fraction from lipid/myelin debris the resulting one cell suspension was further purified with OptiPrep (Alere Technologies AS, Rodelokka, Norway) gradient solutions. Single cell suspensions in HBSS were carefully layered on top of the Optiprep gradient. After centrifugation the top layer constituted the lipid/myelin debris, and underneath were the various cell fractions in distinct layers.

Copyright and License information:  ©2016 Casciati et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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