2. Add 3 ml of STV for T-25 or 5ml for T-75 and discard it to remove floaters.
3. Add 3 ml of STV for T-25 or 5ml for T-75
4. Wait for 2-3 minutes (until the cells peel off from the flask)
5. Add 6ml of DMEM with 10% serum for T-25 or 10ml for T-75
6. Flush the cells in the flask using a sterile pipette
7. Transfer the cells into a sterile falcon.
8. Spin at 1500 rpm for 3 minutes at 27'C
9. Discard the supernatent and suspend the pellet with 3 ml medium
10. Add 6ml of meduim for T-25 or 10ml for T-75
11. Seed 1ml of the cell suspension for T-25 and 2ml for T-75
12. Care is taken to seed the cells uniformly.
13. Avoid formation of clumps.
14. Incubate the flask in incubator at 37'C in 5% CO2