I am infecting human ES cells with a lentiviral construct containing shRNA and am using a non-targeting vecor as control. However, in my non-targeting control I see a high level of differentiation in the cell culture. Is this to be expected? If not are there ways by which I can avoid this.
Transfection or transduction both are stressful to the cells and it is known that some amount of differentiation does occur as a result of stress/passage. However this usually remains background and not more than 5% and hence usually doesn't affect the experiment at large if proper controls are incorporated.
hey buddy
Transduction can induce some gene expression changes like antioxidant defense genes and likely NfKb ,..
In my experience, you may increase your media a little bit when you add virus to the cells or removing a half of media of the cells and replace it with the same amount containing virus to induce the least stress to the cells. you may need to change the media before 24 hours after transduction.
good luck
Subhojit Sen
Hey, do you have an article that proves that transfection or transduction both are stressful to the cells?
Thank you!
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