Dear Daniel,

We always use LipoGeneTM Transfection Reagent, it shows very high transfection efficiency. You can find detailed information in the following website: https://www.genemedi.net/i/lipogene-transfection-reagent

Hope this may help you.

Lentiviral vectors from Addgene. www.addgene.org

Lipofectamine LTX/Plus works well in NIH/3T3 cells (65% GFP transfection efficiency), it will give you higher transfection efficiency in your MEF cells. If you use 24-well plate format to do your experiment, the transfection protocol will be DNA/well : 0.5ug+ 0.5ul Plus in 25ul Opti-MEM; Lipofectamine LTX /well: 1.75ul in 25ul Opti-MEM ; Combine these two solutions, 5min RT incubation, then complex to cells; Post-transfection 48hrs to analyze your experiment.

Please take a look the detail protocol at: http://tools.invitrogen.com/content/sfs/manuals/lipofectamineLTX_and_PLUS_man.pdf

Based on the protocol, you can scale up and down your experiment.

If you are doing siRNA transfection , you might use Lipofectamine RNAiMax as your transfection reagent. The detail protocol will be at:

http://tools.invitrogen.com/content/sfs/manuals/lipofectamineRNAiMAX_man.pdf

Goodluck!

I agree with all. If you dont have a lentiviral system, lipofectamine is the best option.

Thanks for your answers!

I will try Lipofectamine with my MEF cells. If this won't work that well I will check if transduction is possible method in the lab.

I used to use Dharmafect 3 with many cervical cancer cell lines for siRNA transfection. Lipid concentration was 0.05%, I found this concentration optimal with very low cytotoxicity and yet not diluted. Lipofectamine 2000 was the second best option.

Good luck :)

Hi, lipofectamine is only for transient transfection, right? If I want to do stable transfection, is lentivirus the only option? Or is it the best option?

For MEF, I recommend either DreamFect or Magnetofectamine which is a magnetic nanoparticles formulation combined with lipofectamine to increase the efficiency. if you want publications showing that these reagent works well on MEF let me know [email protected]

Thank you for this reccomendation. I will have a look for them.

I tried quite a lot of different reagents and I got the best results using such magnetic nanoparticles (NeuroMag, Nanotherics) or Viromer RED (Lipocalyx).

Both work quite well for the transfection of MEF.

Hi Daniel,

Yes I agree NeuroMag is also pretty good for those cells.

Dear Daniel, you can please follow the links below to obtain certain valuable informations:-

1. http://signagen.com/index.php?main_page=product_info&products_id=768

2. http://ezbiosystems.com/view2.asp?d_id=344

3. https://lifescience.roche.com/wcsstore/RASCatalogAssetStore/Articles/Protocol_X-tremeGENE%20HP_MEF.pdf

Further you can also go though the document attached herewith.

Lipofectamine 3000 is meant to be more efficient than 2000. Follow this link to check it out. http://www.lifetechnologies.com/uk/en/home/brands/product-brand/lipofectamine/lipofectamine-3000.html#resources

Here's a kit for MEF cells (https://altogen.com/product/mef-transfection-reagent-mouse-fibroblast-cells/). It has the transfection reagent and buffers to improve efficiency if you need it. Be sure you know what you're trying to achieve - the question is a bit vague, but if you're doing siRNA transfection high efficiencies should be sufficiently easy to reach.