how to maintain cell in serum free medium - (Jul/19/2007 )

I am going to use siRNA. Can you please help me how I can transfect in to endothelial cells? I like to know whether it is required to maintain cell in serum free medium after transfection of siRNA ? It requires to keep the cell survive more than 4 - 5 days after transfection. But these endothelial cells may not be able to survive 4-5 days in serum free medium. please guide me. [size="3"]

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There are transfection reagents for siRNA, search the forum, you will find it.

usually, the cell are in serum free media after transfection but they are fed with new media with serum after few hrs.

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lipofectin was developped for endothelial cell lines transformation.
Use it with your duplexes will be good.
ttransection is done in medium without serum and without antibiotics.
5h or after 24h you put back the serum on medium.


only if your exp deals with serum starvation :
IMR 90 serum starvation assays are done with 0.2% serum over 48h. At thispoint, the transcription rna polymerase III is alost totally inhibited.
So 5 days..... maybe with 1% serum they gonna survive?
but you need to do an assay with and an assay without serum

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Normally, serum-free medium is required only during transfection. After 6-8 hr of transfection, you can again switch back to normal medium containing serum.

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Thanks for your suggestion

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Thanks for the advice

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during transfection use no serum as especially albumine disturbs; after transfection, at least 4h, you can switch again to serum-containing medium;

if you do not like to use serum at all, you must add some vital components such as STI (selenite-transferrin-insuline), and EGF

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