The cells are transfected first with siRNA, and after 48 hours, I add AngII. After transfection, the confluency goes down, and AngII makes it even worse.
Hello Sana Sana Qayum
The transfection of primary cells may be causing cell death. Primary cells, which are directly isolated from tissues, are generally more delicate and sometimes difficult to transfect compared to established cell lines. They are also inherently more sensitive to transfection than others, making them more prone to cell death.
Moreover, excessive siRNA can lead to cell death, and there is a probability of off-target effects meaning siRNA can unintentionally target other genes besides the intended one, potentially leading to unexpected cell death or other adverse effects.
So, it becomes necessary to optimize on the transfection reagent. You should experiment with different reagents to find the best balance between transfection efficiency and cell viability. Also, optimize the siRNA concentration. You may start with a low siRNA concentration and gradually increase it while monitoring cell viability and transfection efficiency.
Best,
Angll assoc. with cell death due to apotosis or necrosis cause by hiperoxia . Check culture condition , optimize the dose
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