3D Sandwich Cell Culture

  1. Add gel to insert or well using positive displacement pipette
  2. Prepare cells (14 million cells/mL, in 10uL of media per gel)
  3. Pipette cells slowly onto gel
  4. Add a second layer of gel on the top of the deposited cells
  5. Add media on top of gel and in external well
  6. Incubate for 20 minutes at 37 °C, 5 % CO2, humidified incubator
  7. Remove media from on top of gel carefully
  8. Remove media surrounding insert
  9. Repeat steps 4 7 twice (3 media changes over 1 hour)
  10. Culture as usual