I would like to transduce neuronal and astrogilal primary culture cells from mice or rats with an AAV. Do you know a reliable article that allows to answer to these questions : Should I produce an AAV2/6 ? Should I transduce at DIV6 or DIV2 ?
Hi Marie
You could try AAV-DJ, which is the most efficient serotype in infecting in vitro cells in our experience. Paradoxically, AAV-DJ is not good in tissue infection in vivo.
Genemedi is experienced in AAV production, you could find more information and manual files on this website: www.genemedi.net/i/aav-packaging
I used AAV9 to transduce neurons in vivo by direct injection. In our purpose, AAV9 is very effective. Our collaborator Ronald, one coauthor, had several publications that may be helpful. (Mol Ther. 2008 Jan;16(1):89-96).
Hello Boafeng, Thank you very much for your answer. Unfortunatly, transduction profiles of AAVs between in vitro and in vivo can be really different. We also use AAV1 and AAV9 to transduce neurons in vivo, but I am not sure that there are the best for in vitro experiment. I do not know why...maybe in vitro the receptors for AAVs are not mature ?
This paper may help you.
A survey of ex vivo/in vitro transduction efficiency of mammalian primary cells and cell lines with Nine natural adeno-associated virus (AAV1-9) and one engineered adeno-associated virus serotype
http://virologyj.biomedcentral.com/articles/10.1186/1743-422X-10-74
Hi Marie,
Here are some additional references for primary brain cells that should be helpful:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2367141/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2293646/
It seems like different labs come to slightly different conclusions for tropism of different serotypes for specific cell types. The best approach is to try a few serotypes and see what works best for you (if you are packaging your own virus)
Personally I have found a hybrid AAV serotype (AAV-DJ) is highly specific for mouse and rat hippocampal and cortical neurons in vitro. I have also found that AAV2-9 also has strong rates of neuronal transduction, but more glia transduction in comparison to -DJ.
I have not personally used a serotype that transduces glia at high rates, but I am sure there are others who have.
You can also look at this article to get some more information regarding serotypes:
http://www.ncbi.nlm.nih.gov/pubmed/18054899
Good luck
Hi Nicolas, my experiments are not related the hippocampus but you can have a look on this article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4251317/figure/F4/
It shows that, in the hippocampus, the cells transduced by an AAV2/9-CMV-eGFP are 73 % of neurons and 25% glia. ...But...I am not sure you can extrapolate these results because the AAV9 has been injected via the cisterna magna, and not into the parenchyma....and another thing : When I injected an AAV9-CMV-eGFP into the substantia nigra, I did not quantify it , but the transduced cells were around 90% neurons and 5% glia I would say...
Hi Marie
You could try AAV-DJ, which is the most efficient serotype in infecting in vitro cells in our experience. Paradoxically, AAV-DJ is not good in tissue infection in vivo.
Genemedi is experienced in AAV production, you could find more information and manual files on this website: www.genemedi.net/i/aav-packaging
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