We have been trying to transfect Namalwa and JEKO-1 cell lines with an eGFP expressing plasmid and have been having a hard time developing a protocol that results in High transfection rates, High viability and eGFP expression. I have always had trouble with transfecting suspension cells but these seems especially resistant... does anyone have any tips or tricks to these cell lines? We have tried electroporation with Lonza and receive some especially high amount of dead cells and lipofectamine type assays result in very few less then 1% transfection rates... Post sorting of these also did not yield relatively better results.
Hi ondrej,
I am currently also struggeling with transfecting pDNA int Jeko-1 cells.
it would be really nice if you could send me the conditioins of the elctoporation you performed
Ala
Hello,
Lipid based transfection can't be completely ruled out in case of B-cells Lymphoma cell lines. In case of suspension cell lines health of the cells while transfection plays critical role. Normally the chemically defined media used to grow the suspension cell line should have less glutamine supplement . Revival of the electroporated cells usually takes time ( nearly a month)... need to grow the cells with less selection pressure and proper media change . Definitely correct electroporation conditions are important for high efficiency transfection.( Bio-Rad electroporation conditions also works fine for these suspension cell line).
Best regards,
Aparajita
Hi John, the Lonza Nucleofector does work well for the Jeko-1 cells. You just have to make sure that the right program is selected. The Cell Line Kit V in combination with the program X-01 results in 74% transfection efficiency and 50-55% living cells. Similar results were published back in 2006: The proteasome inhibitor bortezomib induces apoptosis in mantle-cell lymphoma through generation of ROS and Noxa activation independent of p53 status.
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