Hello everyone,
I have made a nice protocol for culturing whole adult rat retinas in vitro. This is also called organ cultures or whole mount cultures. The retinas are sitting on a filter in a 12w plate with media in the incubator.
I really wish to perform knockdown using siRNA (I know it works in single cell cultures) on these retinas. I have tried with a lot of lipofectamine and brainfectIN but neither has worked. Lipofectamine normally works great for us on primary single cell cultures, but I think it does not penetrate this many cells deep retina effectively. Most transfection agents are either for single cells or in vivo. In vivo agents may work, as they are used to penetrate tissue more, but so far brainfectIN has not, and not in vivo either.
Does anyone have any experience with effectively transfecting/performing knockdown on organ cultures (penetrating several cell layers) or maybe in vivo? I have found some publication on electroporation but with low efficiency. I need a rather high efficiency to do my experiment.
Thank you!
Dear Hendrik,
It has been a while since you asked the question. Did you manage with the experiment?
You could try magnetofection. There are kits available for in vitro and in vivo transfection - you may need a combination of the two. OzzBioscience has a couple of options available. https://www.ozbiosciences.com/8-in-vivo-delivery-transfection. OzzBio also supply si3Dfect transfection reagent which may fit your application. https://www.ozbiosciences.com/3d-transfection/63-si3d-fect-transfection-reagent-silencing-3d-scaffolds.html. I am sure their tech support will be able to help you.
- Badges
- Science method