Tina:General cell culture protocols: Difference between revisions

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==General ES cell culture==
==General ES cell culture medium==
1. [[mouse ES cell culture medium]]
*'''mouse ESCs'''
1. [[mESCs culture medium]]


2. [[mouse ES cell freezing medium]]
2. [[mESCs freezing medium]]


3. [[human ES cell freezing medium]]
*'''human ESCs'''
3. [[hESCs culture medium]]


4. [[hESCs freezing medium]]




==Feeder dependent ES cell culture==
1. [[Preparation of 0.1% gelatin solution]]


2. [[Preparation of gelatin coated plates]]
==Feeder dependent ESCs culture protocol==
*'''Coating'''
1. [[Preparation of gelatin coated plates]]


3. [[MEF culture medium]]
*'''MEF'''
2. [[MEF culture medium]]


4. [[MEF conditioned medium]]
3. [[Preparation of inactivated MEFs]]


5. [[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 hES cell culture (Matrigel)==
7. [[Passaging hESCs on feeder]]
 
*'''Cryopreservation'''
8. [[Cryopreservation of ESCs]]
 
==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]]


3. [[Passaging hESCs on Matrigel plate (HUES)]]
4. [[Passaging hESCs on Matrigel plate (HUES)]]


4. [[Passaging hESCs on Matrigel plate mechanically (H1, H9)]]
5. [[Passaging hESCs on Matrigel plate mechanically (H1, H9)]]





Latest revision as of 22:27, 7 May 2013

General ES cell culture medium[edit]

  • 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[edit]

  • 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)[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