Jacobs:Protocol Cell Culture of MDCK cells

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MDCK cell culture

JCC 2/25/11

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


  • 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