Hi everyone,
I am about to perform electroporation (Neon) of NK-92 cells with eGFP mRNA. I was wondering why is electroporation efficiency so low in NK cells (40-70%) and survival (~50%) after electroporation? Is there a way to increase both the electroporation efficiency and their survival? Would cells survive better if provided with fatty acid/cholesterol-rich media during or post-electroporation?
Is it holes in cell membrane, ROS and electrolytes that kill cells? If I eliminate all or nearly all parameters that lead to cell death post-electroporation, do you think that NK-92 cells would survive better?
Thanks,
Maria
Dear Maria
I can't help you in optimizing electroporation approach. Nevertheless, you can use home-made lipoplex or lipid nanoparticle to transfer eGFP mRNA into NK-92 cells. Besides, some commercially available reagents such as lipofectamine (as a lipoplex forming mixture) show an acceptable performance in cytoplasmic delivery of mRNAs without significant decrease in the cell viability. However, a bit optimization should be done in each case.
NK-92 is a human natural killer cell line that is commonly used in research and therapeutic applications. However, its survival and electroporation efficiency can be low for several reasons, including:
To increase the survival and electroporation efficiency of NK-92 cells, it is important to optimize the electroporation procedure and to use appropriate quality control measures to ensure that the cells remain viable and functional. Providing the cells with a fatty acid/cholesterol-rich media during or post-electroporation could help to improve their survival, as these compounds have been shown to have a protective effect on cells during electroporation.
It's important to keep in mind that the optimal conditions for increasing the survival and electroporation efficiency of NK-92 cells can vary depending on the specific application and the desired outcome. Further research is needed to determine the best strategies for improving the survival and efficiency of these cells.
I hope this can help. Thank you.
Hi Maria Victorova,
In my experience, the volt and the pulse number have an impact on viability. It requires severe optimization depending on the device you use. In order to increase the efficiency you would consider adding 1.25% DMSO during or after the electroporation. You can check the article below;
https://pubmed.ncbi.nlm.nih.gov/8932394/
Previously, I experienced that the same protocol for one plasmid yields good survival and efficiency results while the other does not. So, you might also check for your plasmid if it is the problem.
Also, I find using fatty acid/cholesterol-rich media logical, and I consider trying. Which additional substance are you considering to use?
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