Hi everybody! I have seen that many people have problems with their primary cultures. In my lab we make primary cultures of rat cortical neurons, and they stay alive for 2 weeks.
First, we use E18-E19, and we disgregate only with the pipette, first with the 1000ul one, after with the 5ul one and finaly with a pasteur pipette which have the end modify by burning it to get a thinest hole. We make it in plating medium: MEM+3G, FBS, HS and B27. We make the dilution and we plate the neurons into a 24 well plate. Three days later we change de medium with new plating, three days later with changing medium (MEM+3G, HS, FUDR+URIDIN). Finally, after three days we change de changing medium by final medium (MEM+3G, HS), and so on.
But I have a question, does anybody know if I can transfect my neurons if I have changed the medium to changing madium the day before? Thanks!
primary culture survival solution and transfection problem
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