Is it possible to use Lipofectamine to transfect plasmid DNA into MCF-7 cells using a 96 well method? Is there a specific protocol you could recommend?

Answer

Yes, Lipofectamine 3000 transfection reagent works well for delivery DNA into MCF-7 cells.

We get ~60% transfection efficiency (GFP plasmid) by using Lipofectamine 3000 to transfect MCF-7 cells. Here are some suggestions for 96-well plate format:

•The day before transfection, seed 25,000 MCF-7 cells per well of a 96-well plate. This translates to ~85% confluence at the time transfection
•Next Morning, transfect the cells (for each transfection reaction):

a)Tube 1: Dilute 100ng DNA ( high quality, endotoxin-free) + 0.2ul of P3000 reagent in 5ul Opti-MEM I
b)Tube 2: Dilute 0.3ul Lipofectamine 3000 reagent in 5ul Opti-MEM I
c)Combine the two tubes and incubate at room temperature for 5 minutes to form transfection complex
d)Following the complex formation step, add 10ul complex directly to the cell culture medium in each well.
e)6 hours post transfection, change cell media and replace with fresh MCF-7 culture medium
f)Post-tfx 48hrs, harvest the cells

The medium change at 6 hours post transfection is not required when using Lipofectamine 3000, but if the cells are sensitive, such as the case with MCF-7, medium changing will help to reduce the cell death.

ATCC MCF-7 cells growth rate is low. We use Gibco DMEM, high glucose, GlutaMax Supplement, pyruvate + 10%FBS to culture the MCF-7. The supplement sodium pyruvate will help MCF-7 growth a lot.