Haynes:MamCultureMedia
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Mammalian Culture Media
"Wild type" stock cells (non-transgenic)
Cell line | Incomplete media | Supplements for complete media (required) | ||||
HEK293 (adherent) | DMEM, high glucose | 10% FBS, 1% pen/strep | ||||
K-562 (suspension) | Iscove's Modified Dulbecco's Medium (IMDM) | 10% FBS, 1% pen/strep | ||||
SK-N-SH (adherent) | Eagle's Minimum Essential Medium (EMEM) | 10% FBS, 1% pen/strep | ||||
U-2 OS (adherent) | McCoy's 5A | 10% FBS, 1% pen/strep | ||||
MCF7 (adherent) | Eagle's Minimum Essential Medium (EMEM) | 10% FBS, 1% pen/strep, 0.01 mg/mL human recombinant insulin | ||||
BT-474 (adherent1) | ATCC Hybri-Care Medium2 | 10% FBS, 1% pen/strep, 1.5 g/L sodium bicarbonate | ||||
BT-549 (adherent) | ATCC RPMI-1640 Medium | 10% FBS, 1% pen/strep, 0.0008 mg/mL human recombinant insulin | - | MCF 10A (adherent) | MEGM BulletKit w/o GA-1000 (gentamycin-amphotericin B mix) | 100 ng/mL cholera toxin3 |
1 BT-474 cells form adherent patches of epithelial-like cells. The patches are compact multilayered colonies that rarely become confluent.
2 Hybri-Care Medium is a powder that needs to be dissolved in water before use.
<sup<3> For best results, do not filter MCF 10A complete medium before use.
Transgenic Cell Selection
Cell Type | Genotype | Transfected Plasmid | Medium | Supplements (required) |
Selection antibiotic(s) |
U2OS (adherent) | Flp-in T-REx | pcDNA5/FRT, or V0200 | McCoy's 5A | 10% FBS-tf, 1 % pen/strep | 15 ug/mL blasticidin, 200 μg/mL hygromycin |
U2OS (adherent) | misc. | MV2 or pcDNA3.1/puro | McCoy's 5A | 10% FBS-tf, 1 % pen/strep | 15 ug/mL blasticidin, 1.0 μg/mL puromycin |
Transgenic Cell Maintenance
Cell Type | Genotype | Medium | Supplements (required) |
Maintenance antibiotic(s) |
U2OS (adherent) | Flp-in T-REx, untransfected | McCoy's 5A | 10% FBS-tf, 1 % pen/strep | 100 ug/mL Zeocin, 15 ug/mL blasticidin |
U2OS (adherent) | Flp-in T-REx, hygro-transfected | McCoy's 5A | 10% FBS-tf, 1 % pen/strep | 15 ug/mL blasticidin, 60 μg/mL hygromycin |
U2OS (adherent) | Flp-in T-REx, puro-transfected | McCoy's 5A | 10% FBS-tf, 1 % pen/strep | 15 ug/mL blasticidin, 0.5 μg/mL puromycin |
HEK 293 (adherent) | Flp-in T-REx | DMEM, high glucose | 10% FBS-tf, 1 % pen/strep | 100 ug/mL Zeocin, 15 ug/mL blasticidin |
HEK 293 (adherent) | luc Gal4-EED "23;4;9" (K. Hansen) | DMEM, high glucose | 10% FBS-tf, 1 % pen/strep | 0.5 ug/mL puromycin |
Notes
- FBS = fetal bovine serum
- FBS-tf = tetracycline-free FBS (required to maintain the "off" state of dox-inducible transgenes)
- pen/strep = penicillin and streptomycin anti fungal supplement
- Filter-sterilize all media before use
- Store all media at 4°C