looking for some assistance.. how do you determine which MCF-7 cancer cells have been well cultured in flasks just by observing the color of the contents in the flask and microscopic view?
The cells were cultured in a mixture media of 90% RPMI 1640, 10% FBS serum (ATTC, Manassas, VA) and 1% Penicillin/Streptomysin (Invitrogen, Carlsbad, CA). The cells were grown in an incubator at 37 °C with 5% CO2.
For electroporation, the cells were washed twice with 1 × PBS, whose pH was adjusted to 7.4, and left in serum-free 199 medium (Invitrogen) for 24 hours. Cells were dissociated from the incubation flask with 0.25% trypsin/EDTA (ATCC) solution. A hemocytometer was used to obtain a final concentration of 1 × 106 cells/mL. Aliquots of 750 μL in 0.4 cm cuvettes were used for electroporation by adding RPMI 1640 medium with 10% charcoal stripped fetal bovine serum.
- Badges
- Science topic