We have infected both primary mouse stem and progenitor cells and primary human stem and progenitor cells with Sigma Aldrich’s VSV-G pseudotyped lentivirus to great success. Does anyone know if it is possible to infect rodent (rat) primary neurons in this manner?
Sure is possible, works great.
Really, you don't need to do anything special either. I don't even isolate or concentrate the virions. I generally just take the virion containing supernatant from the transfected HEK cells and transfer it directly to my cultured neurons (you may need to test a few different volumes to get the right amount infectivity... I use about 50µL of virion containing media per well on a 24-well plate, but your mileage will certainly vary).
Sometimes, getting preferential neuronal tropism (as opposed to glial or other cells) can be a bit of a challenge. The FBS you use, the confluence of your HEK cells upon transfection, and the pH of your media may play a role. This reference may be of some aid:
http://www.nature.com/protocolexchange/protocols/153
also, here is one (of many!) papers using lenti-virus in cultured rat neurons:
http://jcb.rupress.org/content/183/5/893.long
Hi, if you need detailed info you can download this paper recently accepted in gene therapy were we show lentiviral tranduction in vitro and in vivo of motor neurons, DRG neurons, microglia and astroglia.
Cheers
https://www.researchgate.net/publication/230807202_EFFICIENT_GENE_EXPRESSION_FROM_INTEGRATION-DEFICIENT_LENTIVIRAL_VECTORS_IN_THE_SPINAL_CORDPeluffo_Hugo_%28PhD%29*Foster_Edmund_%28PhD%29_Ahmed_Sherif_G._%28PhD%29_Lago_Natalia_%28PhD%29_Hutson_Thomas_H._%28PhD%29_Moon_Lawrence_%28PhD%29_Yip_Ping_%28PhD%29Wanisch_Klaus_%28PhD%29_Caraballo-Miralles_Victor_%28PhD%29_Olmos_Gabriel_%28PhD%29_Llad_Jernia_%28PhD%29_McMahon_Stephen_B._%28PhD%29_and_Yez-Muoz_Rafael_J._%28PhD%29_Accepted_Gene_Therapy
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