- Thaw chondrocytes following the recommended cell-thawing protocol.
- Adjust cell concentration to 0.5–1 million per mL.
- Add to culture flasks or wells at 1–3 x 105 per cm2.
- Incubate at 37°C, 5% CO2.
- Passage cells just before they reach confluence using 0.25% trypsin-EDTA to remove cells from the culture surface.
- Transfer cells to new culture vessels at the same concentration as in Step 3 (1–3 x 105 per cm2). Incubate at 37°C for 1–3 days.
- Continue culture as before.
- We recommend using the cells soon after thawing, within the first three passages.