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Round Bottom Type <Tumor cells>

1. Cell Culture Plate

Micro-Space Cell Culture Plate: Elplasia RB 500 400 NA

500μm(W)

2. Culture Medium

The following medium supplemented with 10% FBS (inactivated, Hyclone) was used for HT-29.

  • HT-29: McCoy's 5A medium (GIBCO®, 16600-082)

3. Cell Seeding

  1. 1000μL, 300μLof culture medium was added in each well of 6- or 24-respectively*.
  2. 2000μL of suspension (5.0x105 cells/mL) was added in each well of 6-well plate. 500μL of suspension (4.0x105 cells/mL) was added in each well of 24-well plate.
  3. The plate was carefully placed into water-jacketed incubator under 5% CO2 and 95% air at 37℃.

4. Medium Change

  1. Half of the medium was replaced by fresh culture medium once or twice a week.
  2. Liquid handling must be performed gently because the spheroid in each compartment tends to detach, float and move.

Important Notice

In case solution contains bubbles when medium placing, they should be removed by pipetting or centrifugation (less than 1000G).

Morphology (2weeks)