Preparation of Condition Media

Materials
  1. Sub-confluent mammalian cells
  2. Culture medium (DMEM+10% FBS + 1% 100X Pen-Strep)
  3. Trypsin-EDTA solution
  4. Sterile PBS
  5. Pipets
  6. Larmini flow hood
  7. CO2 Incubator
  8. 10cm or 6-well plate(s)
  9. Centrifuge
Procedure
  1. Seed cells 1-2 days prior to conditioning at the density that the cells can reach almost confluent.
  2. Wash cells twice with sterile PBS
  3. Add serum-free media (DMEM) at 5ml/plate for 10cm plate, 1ml/well for 6-well plate, or 0.5ml/well for 12-well plate.
  4. Incubate at 37oC with 5% CO2 for 48 hours.
  5. Collect media and centrifuge at top speed(4000rpm) at 4oC for 15 min.
  6. Decant and collect supernatant.
  7. Proceed for analysis or store at -80oC.
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