We are trying to transfect lymphoma cell lines, and we have tried several methods, the best is through the use of lentivirus of course, but in this pandemic days is very difficult to import this kind of vectors. So, I wonder if any have experience with a simpler and even cheaper method??
Hi Lucia, I'd suggest electroporation. I know it can be challenging. However once parameters such as voltage and pulse duration is optimized, you can reach an around 50% tranfection yield with 50%viability if that's adequate for your experiment.
Electroporation offers a high transfection efficacy but on the other side many cells die in this setting (up to >90% depending on the cell line).
Even in suspension, lipofection is an adequate method which leads to an adequate transfection rate (depending on the cell line) associated with a lower degree of cell death in your setting.
If you just need to obtain a recombinant protein quickly, PEI-mediated transient transfection can be great (and very cheap). If you need stable transfected cell lines/clones then you should analyze other variants as described.
Thanks for your recommendations, we have tried with lipofectamine an it didn´t work, but we will try with electroporation. Maybe the problems we have had are specific to the lymphoma cell lines we are using
As you asked for the simpler and cheapest method as an alternative for transfection in a suspension cell line, the following attached article can help you to try something new.
Hope it might help.
you can try nucleofection, they optimized protocol for each cell type, you will find your cells of interest in their database. may be wait for few days to get restrictions of COVID19 over.
I would recommend the Nucleofector technology. We have over 700 transfection protocols for hard to transfect cells like lymphoma cell lines, but also primary lymphoma and leukemia cells. For around 400 cells we have the ready-to-use protocols where not optimization is needed, just select the cell type from the pre-installed list and start with the real experiments. Do not spend time on optimization but on real research. All protocols and transfection data can be found here: https://knowledge.lonza.com/search-results?search=*&type=Cell%20Information&orderby=relevance
Dear Peter thans so much for your recommendation I will check that system carefully.
Try PEI based transient transfection. It is one of the cheapest way to do it. Go for the ratio of 3:1 that works for the best. I have tried that with my suspension 293T cells and it works the best. I would only suggest electroporation for stable transfection.
Dear Rupsa,
we have tried with PEI and lipofectamine and didn´r work well , but we will ry with electroporation. Thanks¡¡
You can try Neon transfection as well. They have protocols for BC-1 and other cell lines, check their cell line database. You can get 20-30% efficiency on some hard to transfect cell lines.
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