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Square Type <Tumor cells>

1. Cell Culture Plate

Micro-Space Cell Culture Plate: Elplasia SQ 200 100 NA


2. Culture Medium

The following medium supplemented with 10% FBS (inactivated, Hyclone) was used for A549, HT-29, MCF-7, NCI-H640 and T47D. Only SKOV3 cells were grown in the following medium with 15% FBS.

  • A549: F-12K (GIBCO®, 21127-022)
  • HT-29: McCoy’s 5A medium (GIBCO®, 16600-082)
  • MCF-7: EMEM medium (Wako, 051-07615)
  • NCI-H640: RPMI1640 (GIBCO®, 22400-089)
  • SKOV3: McCoy’s 5A medium (GIBCO®, 16600-082)

3. Cell Seeding

  1. 300μL, 50μL or 15μL of culture medium was added in each well of 24-, 96- or 384-well plate, respectively.
  2. 500μL of suspension (4.0x105 cells/mL) was added in each well of 24-well plate. 100μL or 33μL of cell suspension (3.3x105 cells/mL) was added in each well of 96- or 384-well plate, respectively.
  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).


Cell line Culture period
1 Day 3 Day 6 Day

Medium Change 200μm