I am working with L6 CELL line. I am trying to transfect them using lipofectamine 2000. But none of the cells are getting transfected, even though the same protocol is working for other cell types. please help.
Lipofectamine 2000 may work well for some cell lines but not for others. Have you ever thought about trying different transfection reagents such as L6 Transfection Reagent (#6837, Altogen Biosystems) and DNAfectinTM 2100 (G2100, Applied Biological Materials Inc.)?
Good luck!
Dear Priyankar,
L6 cells can be considered hard-to-transfect cells and it's quite true that chemicals and classic transfection reagents show some limits.
That's why I would highly recommend you to try Magnetofection method that uses a Magnetic field to concentrate complexes of magnetic nanoparticles/nucleic acids onto the cell surface.
Even if it is considered as a physical way of transfecting cells, the Magnetofection uses the classic pathways to transfect cells (endocytosis...); no hole is created during the transfection process and cellular integrity remains intact.
All the material are biodegradable and compatible with cell and organisms life.
This methods is really efficient for primary cells and hard-to-transfect cells ; the best example is be the transfection of primary neurons, primary endothelial cells, primary amniotic cells...
you can find more information at:
http://www.ozbiosciences.com/
the best solution within the reagent would be PolyMag Neo (http://www.ozbiosciences.com/polymag-neo.html) and the Magnetofectamine Kit that associates Magnetofection to Lipofectamine 2000 (http://www.ozbiosciences.com/transfection-dna/49-magnetofectamine-magnetofection-lipofectamine2000-combimag.html).
you can contact me directly at: [email protected] for more information,
I hope this will help!
good luck with your experiment,
Cedric
http://www.ozbiosciences.com/
respected Cédric Sapet
we use neuromah from oz biosciences. as per your suggestion i will try to use magnetofectamine kit. I want to ask whether neuromag can be used for transfecting l6 cells, as I have the reagent with me?
thank you for the suggestion
Dear Priyankar,
For now, we do not have feedback from collaborators that could have use NeuroMag for L6 cells. First dedicated to primary neurons transfection, numerous articles demonstrated that it can also work on neuron cell types (from astrocytes to neural stem cells) and also on several cell lines.
So as you already use NeuroMag, you can try it on your cells ; I would recommend :
- use a fixed DNA amount and vary NeuroMag ratio
- once the correct ratio is found, keep it unchanged and vary the DNA amount.
This should lead to optimized transfection conditions.
I hope this will be of help,
do not hesitate to contact me for any question,
best regards,
Cedric
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