Hi, we have been doing cortical neuron culture from E-17-18 Embryos of rat and so far all has been good but all of sudden we are having a strange phenomena that after 3 days or sometimes 2 days of division they are dying. earlier we kept them for 5 days of division and they remained healthy. We change media every alternate days and once this happened we checked all our reagents including B27 and FBS, but we are unable to figure out what's causing this problem as it is hampering our work badly. Any suggestion will be kind help.
Hello Disha,
I had this problem long time ago, and when discussing with oter persons who realize primary neuron culture, it seems that B27 was defectious (Invitogen sent us a new batch). Be careful with CO2 too, and look if you have enough water in th incubator.
Good luck
Frédérique
Thank you Frederique, but we already tried B27 from other lab and still neurons are dying,we already checked incubators they are good
Dear Disha,
I would check if everything is fine with your coating i.e. pH of H2O you use to dilute your PDL and etc.
Good luck!
Natalia
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