Hi community,
At the moment, I transfecting primary ventral horn mixed motoneuron cultures from E13 mouse embryos with small RNA fragments (30-45nt) to see their properties in protection against different stress. I transfected ~750nM RNA fragments with Lipofectamine2000 for 1h and stress the cells for 24h. Problem is, ~60% of the motoneurons (SMI32 positv counts by ICC) are dying due to transfection. Cell death of stressed transfected MN is as high as without stressing transfected MN.
I compared transfection efficiency between RNAiMAX, FuGENE HD, Lipofectamine2000 and Viromer Blue, and Lipofectamine showed the best transfection.
What reagent would you recommend to try?
Thanks
Hi Sergej,
I remember in my last lab we were trying different transfection reagents for both toxicity and efficiency, and the Mirus Transit-X2 reagent found to be best. It even outperform Lipofectamine. I never tried it on primary cells, but on cancer cells it was more efficient and you need to use less of the reagent which makes it less toxic. I think you can get free trials from them sent to your lab. Also you might think of giving your cells a bit more time after transfection before you put them under stress, of course if your experimental procedure allows. Here is the link to Transit-X2.
Best of luck,
https://www.mirusbio.com/products/transfection/transit-x2-dynamic-delivery-system?gclid=CjwKEAiAmeyxBRCJxoKk7IWLl2oSJABvZjhhJJM-mUZfnUITusE1E4_OgsaXbOfp5lHwrQaX09JoGBoCVRjw_wcB
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