Hi
I would like to transiently transfect SHSY5Y cells. I know they are not very easy transfect and I read one article where they used Lipofectamine 2000 but on invitrogen protocol pages I did not see any information for SHSY5Y cell transfection. anybody has an experience? and maybe protocol?
You should look into transfection reagents that are pre-optimized for cell lines like SH-SY5Y (https://altogen.com/product/sk-n-sh-transfection-reagent-neuroblastoma-cells-htb-11/). As long as you follow designated protocols, you should be fine, but coming up with one on your own is an investment that could be avoided easily. Nonetheless, SH-SY5Y can be transfected with high efficiency (lower for plasmids) so you should get good results for the most part.
I have experience with shy and sk-y cells. I had good results using freshly passed cells (the night before) and Jet-PEI polyplus transfection reagent, which has the advantage of being serum and antibiotics compatible. Usually the efficiency is around 30-50% (sometimes higher) which is suitable for most biochemical applications.
In my experience reverse transfection with RNAiMAX works best for siRNA, and nucleofection protocol g-004 works best for plasmids.
For RNAi, 25pmol in a 6well with 250k cells gave 70% KD after 24hr.
For plasmid transfection, 1ug of pmaxGFP in 1M cells gave up to 80% efficiency.
Dear Sleman,
I would propose you an alternative to L2000 that was demonstrated by several publications to work very well on SH-SY5Y.
DreamFect and DreamFect Gold are two transfection reagents that are really efficient for many cell types, of whom SH-SY5Y, you can refer to the following references:
Donnelly PS., Proc Natl Acad Sci U S A. 2012 Jan 3;109(1):47-52.
Grohmann M., BMC Cancer. 2009 Aug 27;9:301.
Parker SJ., PLoS One. 2012;7(8):e42277.
Shao M., Nucleic Acids Res. 2011 May;39(10):4035-47.
Shao M., Nucleic Acids Res. 2011 May;39(10):4035-47.
Shao M., Nucleic Acids Res. 2011 May;39(10):4035-47.
Krol HA., PLoS One. 2008 Jan 30;3(1):e1503.
do not hesitate to contact me if yo uwould like to try these reagents, (you can also contact me directly at: [email protected])
good luck with your experiments,
best regards,
Cedric
Thank you both Martin Forde and Cédric Sapet
Cédric this week i would to transfect my cells with JETPEI if it dosn't work i will contact you...may you have cheaper L2000 than another companies.. :)
BEST
It looks like my suggestion will be kind of late for you, but for the other people I highly suggest nucleofection. I'm using Amaxa Kit V, and it works really well.
You should look into transfection reagents that are pre-optimized for cell lines like SH-SY5Y (https://altogen.com/product/sk-n-sh-transfection-reagent-neuroblastoma-cells-htb-11/). As long as you follow designated protocols, you should be fine, but coming up with one on your own is an investment that could be avoided easily. Nonetheless, SH-SY5Y can be transfected with high efficiency (lower for plasmids) so you should get good results for the most part.
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