Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/09/17

Growing Up Cells

The previous attempt to get the cells started officially did not work. The cells did not attach and were floating, even after three days of incubation at 37C. I did another thaw and this time I did the following:

1. Added thawed cells to 5 mL pure FBS in 15 mL conical tubes
2. Spun at 1000 rpm for 4 minutes
3. Aspirated off the FBS (to get rid of any toxins from the freezing solution)
4. Resuspended the cell pellet in 1mL 20% FBS/ 1% pen-strep DMEM media
5. Added the resuspended cells to 3 mL of 20% FBS/ 1% pen-strep DMEM media in a 6-well culture dish (one cell line per well). This gives the cells less surface area to stick to and crowds them together, which sometimes promotes cell growth.

The cell lines I thawed are

1. HepG2
2. Luc #4
3. 23;4;9 Luc EED