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Dissecting neonatal brains to get glial cells

Today in the lab I was dissecting neonatal brains, in order to get glial cells and culture them for my experiments. I dissected a total of 7 brains, which took me about 6.5 hours without any breaks. The process is relatively straight-forward, but it requires a lot of dexterity because the brains are very small and delicate.

Neonatal brains that I dissected for today's experiment.
Neonatal brains that I dissected for today's experiment.

Detailed methods of today’s experiment

The hippocampi and cortices were dissected out of each brain and minced using spring scissors. The tissue from each brain was transferred in cold dissection medium and trypsin was added to dissociate the cells. The cells were incubated in a 37°C water bath for 15 minutes, swirling frequently. Then, trypsin inhibitor was added to inhibit trypsin activity, and the tube was left to incubate for 1 minute at room temperature. After that, DNase I was added to digest the sticky DNA released from dead cells, and the tube was centrifuged at 400 × g for 5 minutes. The supernatant was removed, the pellet was triturated in pre-warmed culture medium and centrifuged again at 400 × g for 5 minutes. The supernatant was removed, and the pellet was resuspended in 5 ml pre-warmed culture medium. The cell suspension was cultured in a PDL-coated T-75 tissue culture flask with pre-warmed culture medium and incubated at 37°C, 5% CO2.