Passaging hESCs on feeder

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1. Aspirate media from cells

2. Wash well once with PBS

3. Add 1ml TrpLE to each well, incubate 5 min at 37C

4. Remove TrpLE

5. Add 1ml H9 basal media and detach by scraping

6. Pipette using p1000 2-3 times to dissociate

7. Transfer 100ul (1:10) to 50ul (1:20) dissociated cells to fresh feeders (3ml total media + bFGF and Y27632)