Haynes:MamCultureMedia: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 18: | Line 18: | ||
| U-2 OS (adherent) || McCoy's 5A || 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 | | MCF7 (adherent) || Eagle's Minimum Essential Medium (EMEM) || 10% FBS, 1% pen/strep, 0.01 g/L human recombinant insulin | ||
|- | |||
| BT-474 (adherent<sup>1</sup>) || ATCC Hybri-Care Medium || 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 g/L human recombinant insulin | |||
|} | |} | ||
<sup>1</sup> The cells form adherent patches of epithelial-like cells. The patches are compact multilayered colonies that rarely become confluent | |||
<br> | <br> |
Revision as of 12:40, 15 June 2015
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 g/L human recombinant insulin | ||||
BT-474 (adherent1) | ATCC Hybri-Care Medium | 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 g/L human recombinant insulin |
1 The cells form adherent patches of epithelial-like cells. The patches are compact multilayered colonies that rarely become confluent
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