Media for culturing cells
Rebecca Berrens
Published: 2022-08-17 DOI: 10.17504/protocols.io.n2bvj652xlk5/v1
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Abstract
common ES cell media for mouse embryonic stem cells
Steps
ES cell media
1. - some people remove 50 ml of DMEM and replace it with 50 ml of Analar water or just add 50 ml of Analar water to 500 ml of media, but we don’t do this. - fetal calf serum should be batch tested for ES cell growth/differentiation and toxicity
ES Medium| A | B | C | D | | --- | --- | --- | --- | | Reagent | Volume | Supplier | Cat.number | | DMEM without sodium pyruvate* | 500 ml | Invitrogen | 41965-039 | | Fetal calf serum ** (13% final) | 75 ml | GlobePharm | | | Non-essential amino acids, 100x | 5 ml | Invitrogen | 11140-035 | | L-glutamine, 200mM, (100x) | 5 ml | Invitrogen | 25030-024 | | b-mercaptoethanol, 50 mM | 0.6 ml | Invitrogen | 31350-010 | | Penicillin/Streptomycin | 5 ml | Invitrogen | 15140-122 | | LIF (ESGRO) (now we use home-made LIF-conditioned medium) | 1ml (check current titer and add accordingly) | | |
Note
EC10 media
2.
Medium for differentiated ES cells and most fibroblast cultures (EC10)
| A | B | C | D |
|---|---|---|---|
| Reagent | Volume | Supplier | Cat.number |
| DMEM without sodium pyruvate* | 500 ml | Invitrogen | 41965-039 |
| Fetal calf serum** | 50 ml | GlobePharm | |
| Non-essential amino acids, 100x (optional) | 5 ml | Invitrogen | 11140-035 |
| L-glutamine, 200mM, (100x) | 5 ml | Invitrogen | 25030-024 |
| b-mercaptoethanol, 50 mM | 0.5 ml | Invitrogen | 31350-010 |
| Penicillin/Streptomycin | 5 ml | Invitrogen | 15140-122 |
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