I had obtained the DA primary cell culture from P0 mice and would like to test them with my construct via AAV induction. Many electrophysiology experiments were done on the primary cell culture, but seldom with AAV. Some detail has been left out from most papers as after how many days did they wait for their first treatment. Also, how long does the AAV take in effect in primary neurons. Thank you for your input!
Hi,
The reason recombinant AAV (rAAV) is not often use in cell culture is, unlike in vivo, it has low efficiency and it tends to be diluted through cell divisions. rAAV do not integrate and persists in episomic form almost exclusively. This is fine in neuronal primary cell culture since neurons don't divide. However, it will take at least two weeks (depending on the expression cassette and the promoter) to reach optimal expression. I faced this problem in the past and end up using lentivirus, Their optimal expression is within a week and since they integrate, there is no dilution of signal. They also express large transgenes, almost twice that of AAV.
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