Tina:General cell culture protocols: Difference between revisions
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==General ES cell culture== | ==General ES cell culture medium== | ||
1. [[ | *'''mouse ESCs''' | ||
1. [[mESCs culture medium]] | |||
2. [[ | 2. [[mESCs freezing medium]] | ||
3. [[ | *'''human ESCs''' | ||
3. [[hESCs culture medium]] | |||
4. [[hESCs freezing medium]] | |||
==Feeder dependent ESCs culture protocol== | |||
*'''Coating''' | |||
1. [[Preparation of gelatin coated plates]] | |||
*'''MEF''' | |||
2. [[MEF culture medium]] | |||
3. [[Preparation of inactivated MEFs]] | |||
*'''mouse ESCs''' | |||
4. [[Thawing mESCs on feeder]] | |||
5. [[Passaging mESCs on feeder]] | |||
*'''human ESCs''' | |||
6. [[Thawing hESCs on feeder]] | |||
==Feeder independent | 5. [[Passaging hESCs on feeder]] | ||
==Feeder independent ESCs culture protocol(Matrigel)== | |||
1. [[Preparation of Matrigel plates]] | 1. [[Preparation of Matrigel plates]] | ||
2. [[Thawing hESCs on Matrigel plate]] | 2. [[Preparation of MEF conditioned medium]] | ||
3. [[Thawing hESCs on Matrigel plate]] | |||
4. [[Passaging hESCs on Matrigel plate (HUES)]] | |||
5. [[Passaging hESCs on Matrigel plate mechanically (H1, H9)]] | |||
Revision as of 23:55, 26 April 2013
General ES cell culture medium
- mouse ESCs
- human ESCs
Feeder dependent ESCs culture protocol
- Coating
1. Preparation of gelatin coated plates
- MEF
3. Preparation of inactivated MEFs
- mouse ESCs
- human ESCs
Feeder independent ESCs culture protocol(Matrigel)
1. Preparation of Matrigel plates
2. Preparation of MEF conditioned medium
3. Thawing hESCs on Matrigel plate
4. Passaging hESCs on Matrigel plate (HUES)
5. Passaging hESCs on Matrigel plate mechanically (H1, H9)
Osteogenic differentiation media
MEM containing 0-20% heat-inactivated ES-FBS (to be determined), 10−7 M Dexamethasone, 50 μg/ml Ascorbic acid and 10 mM β-glycerolphosphate