siRNA transfection reagent for primary cultured cells - (Nov/25/2005 )

Hi,

I'm going to try RNAi on primary tendon fibroblast cultures. Any advice on the transfection reagents to use for primary cells? I realise the transfection efficency will be low - this is a preliminary expt to determine the most efficienct siRNA sequence before moving onto viral transfections.

---

In my lab, different people prefer different reagents. We have experiences on siRNA transfection reagents like siPORT (Ambion), HiPerfect (Qiagen), X-treme GENE (Roche), RNAicarrier (Epoch Biolabs) and TransPass (NEB)...If you hit the right sequence, the silence effect is quite similar for all reagents. If your sequence is less than perfect, then tansfection efficiency and IFN effect could play a key role in your outcome.

Personally, I like RNAicarrier simply because it is the cheapest, reliable and consistent in my hands on primary MEF (mouse embryonic fibroblsts), primary human keratinocytes and primary rat aortic smooth muscle cells.

---

I'm trying to transfect primary astroglial cells (from P0 mice) with siRNA.

Could anyone suggest a transfection reagent that works for this cell type and a protocol?

Do you know, how many days old cells could be efficiently (more than 80%) transfected?

---