Aug 17, 2022
Media for culturing cells
- 1IDRM, Paediatrics, Oxford University
Protocol Citation: Rebecca Berrens 2022. Media for culturing cells. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvj652xlk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 17, 2022
Last Modified: August 17, 2022
Protocol Integer ID: 68787
Abstract
common ES cell media for mouse embryonic stem cells
ES cell media
ES cell media
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
- 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
EC10 media
EC10 media
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 |