We are interested in AML cell transfections (respectively MOLM14 and MV4-11) by miRNAs or siRNAs. We tried several electroporation or amaxa protocols , but transfection efficiencies were very low or mortality was very high.
Would someone have successfully transfected these cell lines and if so, what was the method and the protocol used ?
Thank you very much for your answers.
For reference see: Onco Targets Ther. 2013 Aug 26;6:1153-60. doi: 10.2147/OTT.S45376.
You can get more than 70% knockdown of MLL/AF4 in MV4-11 if you electroporate 200 nM siRNA in any volume between 50 to 700 µl of (1-10) million/µl cell suspension in normal culturing media, using 4 mm gap cuvette, and 330 V square wave delivered in one pulse for 10 ms. For 2 mm cuvette, just divide the voltage by 2. I would assume most haematopoietic cell lines can be transfected with 300-400 V and 10 ms or 2 pulses of 5 ms. The electroporation buffer is just occult recipe because the electroporators do not provide proper uniform electric pulses.
I agree. Pro: electroporation is quite more efficient if you have enough cells, since Con: death rate is much higher using this methode.
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