Tina:General cell culture protocols: Difference between revisions
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>TinaLo (Created page with '==Feeder independent hES cells culture== ===Preparing Matrigel Solution=== 1. Thaw a stock Matrigel (5 ml/bottle) overnight (at 4°C). 2. Add 5 ml of DMEM/F12 (at 4°C) to the …') |
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== | ==General ES cell culture medium== | ||
*'''mouse ESCs''' | |||
1. [[mESCs culture medium]] | |||
2. [[mESCs freezing medium]] | |||
*'''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]] | |||
7. [[Passaging hESCs on feeder]] | |||
*'''Cryopreservation''' | |||
8. [[Cryopreservation of 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 | |||
== | |||
Latest revision as of 22:27, 7 May 2013
General ES cell culture medium[edit]
- mouse ESCs
- human ESCs
Feeder dependent ESCs culture protocol[edit]
- Coating
1. Preparation of gelatin coated plates
- MEF
3. Preparation of inactivated MEFs
- mouse ESCs
- human ESCs
- Cryopreservation
Feeder independent ESCs culture protocol(Matrigel)[edit]
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[edit]
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