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