I would like to infect a high proportion of mouse cortical neuronal cultures from P0 pups with AAV for a biochemical study while avoiding toxicity. I would like some suggestions on the MOI (ratio of viral genome copies per cell) to use. I am using a AAV-DJ purified via iodixanol ultracentrifugation. I have cells plated in 6 well plates at a density of 10^6 cells/well and in 24 well plates at 150,000/well.
I know there are at least a handful of publications that have made comparisons of different MOIs (see below). The best would be to make my own empirical determination, but wanted to see if anyone had any input, as the some of the publications seem to suggest pretty large ranges in appropriate MOIs( ie 2*10^4 versus 1*10^7). Thanks!
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2367141/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2293646/
Those MOI recommendations are bizarrely large and suggest poor understanding of the concept, low quality viral preparation or transductions in huge volumes. The reason this has to be done empirically is because you have no idea just how good your viruses are or the transduction conditions. I think by definition (using the equation) an MOI of 8 should give you ~100% transfection (assuming perfect conditions).
I am also interested to know what is the good MOI to transfect neurons.
Checking other publications:Article Viral-Mediated Gene Transfer to Mouse Primary Neural Progenitor Cells
Article A survey of ex vivo/in vitro transduction efficiency of mamm...
Hughes, et.al. found that AAV at MOI 2 had no successful transfection. Since then high level MOI was adopted (10^5).
Andrew Coleman, what MOI you end up using?
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