Changing Media for Lentiviral Infection in a Suspension Culture - (Dec/17/2008 )
Hi,
I've used adherent cells for a while now, but am just starting to use suspension cells. As part of my project, I need to infect my suspension cells with lentivrus and then select using puromycin. This process requires changing the media on day 2 to remove the polybrene in the transfection and then again every 2 days after that for 10-14 days to do the puromycin selection. How do I change the media with these suspension cells?
I should also add that these experiments are typically done in 12 or 24-well plates.
Thanks!
-neuroJ-
QUOTE (neuroJ @ Dec 17 2008, 04:54 PM)
Hi,
I've used adherent cells for a while now, but am just starting to use suspension cells. As part of my project, I need to infect my suspension cells with lentivrus and then select using puromycin. This process requires changing the media on day 2 to remove the polybrene in the transfection and then again every 2 days after that for 10-14 days to do the puromycin selection. How do I change the media with these suspension cells?
I should also add that these experiments are typically done in 12 or 24-well plates.
Thanks!
I've used adherent cells for a while now, but am just starting to use suspension cells. As part of my project, I need to infect my suspension cells with lentivrus and then select using puromycin. This process requires changing the media on day 2 to remove the polybrene in the transfection and then again every 2 days after that for 10-14 days to do the puromycin selection. How do I change the media with these suspension cells?
I should also add that these experiments are typically done in 12 or 24-well plates.
Thanks!
Hi
you can put your suspension cell in a sterile Falcon tube and spin down about 2.000 RPM for 10 minutes (do not exceed the rotation because you may damage the cells).
Throw the supernatant and resuspend the pellet with new media.
Put iin new flasks. Remember use everything sterile.
Regards!
-Reis V.P-