This week was very similar to the previous week where the entire week was centered around one experiment. I did mini assays alone throughout the week but the main focus was on the fibroblast cell lines being tested.
This week, as usual, I did a cancer cell assay. I repeated an experiment that I had done a few weeks ago with MCF7, only I increased the growing period from 24 hours to 48 hours. I had previously thought that with such high densities 48 hours would kill off all the cells. However, after getting such low OD numbers (highest OD was 0.63), I decided to test 48 hours. An ideal OD in my experiment should be above 1.5, as I am trying to get maximum cell growth. When I carried out the assay after 48 hours, the highest OD number was 2.3 for the Crystal Violet assay (for MTT it was 0.7 but MTT is always lower than CV).
The fibroblast cells this week were used to point in which regular cells stopped using glucose as energy and switched to ketones. As with the cancer cells the week prior, the fibroblast cells were incubated for 96 hours and checked daily. When the cells switched to a ketogenic source of energy, the concentration of ketones in the media would begin to increase. We monitored and then graphed the growth and energy consumption rate of the cells using mitotract, a dye which traces activity in the mitochondria. When cells stopped using glucose as a source of fuel, mitochondrial activity with severely decrease.