Shreffler:Notebook/retinoic acid trans signaling/2009/05/15

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Growing a T-Cell Line: Jurkat Cells

Jurkat Cells: Tower 6, Box 1 in liquid nitrogen (Lab 17-46), Box Labeled: T6B1, #61 - 1 x 10^7 (9|5 possible strain), dated 10/10/2006

- Obtain cells from liquid nitrogen, thaw them out in hot water bath until a small pellet of ice remains in the cryovial - With a transfer pipet, transfer cells into 50 ml tube and slowly add 10 mls of PBS to allow the DMSO to be diluted - Spin cells at 300g for 10 minutes - Make a medium containing RMPI and 10% Fetal Bovine Calf Serum - Use a 25 cm^3 cell culture flask instead of a larger flask (since there are not too many cells, keeping them together will better ensure their survival) - Resuspend the cells in 1 ml of the RPMI medium - Add 5 mls of the RMPI medium to the culture flask, then add the cells with a transfer pipet - Keep the flask upright with the lid slightly open (these cells are suspension cells, therefore they will not stick)

Prepare an Agar Gel Plate:

- From a pre-made Agar gel solution, take out a 100 mls (this should make 4 to 5 plates) - Bring the solution to a slight boil and add 50 uls of carbinicillin (an antibiotic) - Allow to cool and pour into each plate (covering the complete plate) - Let it sit for 5 to 6 hours at room temperature, then place at 4 degrees to completely dry overnight - Plates must be turned upside-down to ensure no moisture or condensation affects the gel