Passaging hESCs on feeder

From ZhangLabWiki
Revision as of 22:19, 7 May 2013 by >TinaLo (Created page with "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 scrapi...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

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)