Astrocyte Isolation: A Method to Obtain Pure Preparation of Mouse Cortical Astrocytes

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Samenvatting

Source: Schildge, S. et al. Isolation and Culture of Mouse Cortical Astrocytes. J. Vis. Exp. (2013)

In this video, we describe the method to obtain a pure preparation of astrocytes from newly born mice.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Isolation and Plating of Mixed Cortical Cells

Mixed cortical cell isolation for astrocyte cultures can be performed using P1 to P4 mouse pups. In order to achieve proper astrocyte density it is necessary to use 4 mouse pup cortices per T75 tissue culture flask. Therefore, volumes in the following protocol are calculated for a cell preparation using 4 mouse pups.

Before starting the dissection procedure, prewarm 30 mL of astrocyte culture m....

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Resultaten

Brain injury assessment; series of rodent brain images; post-trauma analysis; histological study.
Figure 1. Dissection of postnatal (P3) mouse cortex. A) Whole brain. B) Brain after removal of olfactory bulbs and cerebellum. C) Isolation of cortices by peeling off the plate-like structure of the cortex from the brain. D, D') Cortex from ventral and dorsal.......