Jacobs:Protocol Cell Culture of MDCK cells

MDCK cell culture

JCC 2/25/11

From James Nelson’s lab, cells from Lance Kam's lab

Materials

• Dulbecco's Modified Eagle Medium, low glucose, with L-glutamine, with 110mg/L sodium pyruvate, without sodium bicarbonate. (Sigma D5523-10L)
• Sodium bicarbonate (Sigma S5761-500G)
• HCl
• Filter
• MilliQ water
• FBS
• Pen/Strep

DMEM prep

1. Add 5.0 g DMEM to 475 ml MilliQ water.
2. Stir to dissolve
3. Add 0.5 g of sodium bicarbonate.
4. pH of media should be 7.0 - use concentrated HCl to adjust
5. Bring up to final volume of 500 ml.
6. Filter sterilize with a 0.22 μm filter.

Working media

500 ml DMEM (serum-free) 50 ml FBS 5 ml Pen/Strep

• Grown on TCPS, use collagen coating when grown on glass.

• History: JCC, last updated 2/25/11