I need to mix human neurons with mouse neurons or astrocytes. So I have to pass and count human neurons, but I can immage that is very dangerous and they tend to die. Let me know if somebody has set up a protocol. Thanks
Hi Clara,
I have found that I can passage iPS-derived neurons only within the first two weeks of differentiation. Any later, and all the neurons will die.
I use Accutase for the passage, and leave it on the cells up to 20 or 25 minutes.
Neurons are quite robust and can be replated by trypsinization and centrifugation just as you would passage any other cells. After adding trypsin you have to monitor every 3 minutes that the cells and neurites are detached before removing for centrifugation. Not more than 6 -8 minutes max. Neurons being postmitotic terminally differentiated calls will not divide, but they do regenerate.
Thanks guy. I tried with my neurons at 10 days of differentiation, I used acutase for 3 min and I put also rock inhibitor....everything work! I hope that they will survive during the time.
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