Haynes:MamCultureMedia: Difference between revisions
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| HEK293 (adherent) || DMEM, high glucose || 10% FBS, 1% pen/strep | | HEK293 (adherent) || DMEM, high glucose || 10% FBS, 1% pen/strep | ||
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| K-562 (suspension) || Iscove's Modified Dulbecco's Medium (IMDM) || 10% FBS, 1% pen/strep | | [http://atcc.org/Products/All/CCL-243.aspx#culturemethod K-562] (suspension) || Iscove's Modified Dulbecco's Medium (IMDM) || 10% FBS, 1% pen/strep | ||
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| SK-N-SH (adherent) || Eagle's Minimum Essential Medium (EMEM) || 10% FBS, 1% pen/strep | | [http://atcc.org/Products/All/HTB-11.aspx#culturemethod 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 | | [http://atcc.org/Products/All/HTB-96.aspx#culturemethod 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 | | [http://atcc.org/Products/All/HTB-22.aspx#culturemethod MCF7] (adherent) || Eagle's Minimum Essential Medium (EMEM) || 10% FBS, 1% pen/strep, 0.01 mg/mL human recombinant insulin | ||
|- | |- | ||
| BT-474 (adherent<sup>1</sup>) || ATCC Hybri-Care Medium<sup>2</sup> || 10% FBS, 1% pen/strep, 1.5 g/L sodium bicarbonate | | [http://atcc.org/Products/All/HTB-20.aspx#culturemethod BT-474] (adherent<sup>1</sup>) || ATCC Hybri-Care Medium<sup>2</sup> || 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 | | [http://atcc.org/Products/All/HTB-122.aspx#culturemethod BT-549] (adherent) || ATCC RPMI-1640 Medium || 10% FBS, 1% pen/strep, 0.0008 mg/mL human recombinant insulin | ||
|- | |||
| [http://atcc.org/Products/All/CRL-10317.aspx#culturemethod MCF 10A] (adherent) || MEGM BulletKit without GA-1000 || 100 ng/mL cholera toxin<sup>3</sup> | |||
|} | |} | ||
<sup>1</sup> BT-474 cells form adherent patches of epithelial-like cells. The patches are compact multilayered colonies that rarely become confluent.<br> | <sup>1</sup> BT-474 cells form adherent patches of epithelial-like cells. The patches are compact multilayered colonies that rarely become confluent.<br> | ||
<sup>2</sup> Hybri-Care Medium is a powder that needs to be dissolved in water before use. | <sup>2</sup> Hybri-Care Medium is a powder that needs to be dissolved in water before use.<br> | ||
<sup>3</sup> For best results, do not filter MCF 10A complete medium before use. | |||
<br> | <br> | ||
| Line 71: | Line 74: | ||
</div> | </div> | ||
==BT-549== | |||
Confluent T-75 flask of BT-549 cells, density ~430,000 cells/mL | |||
[[Image:11-23-2015_BT549_confluent.tif | 500px]] | |||
Revision as of 15:56, 23 November 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 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 without GA-1000 | 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.
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
BT-549
Confluent T-75 flask of BT-549 cells, density ~430,000 cells/mL
