Haynes:MamCultureMedia: Difference between revisions
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| '''Cell line''' || '''Incomplete media''' || '''Supplements for complete media (required)''' | | '''Cell line''' || '''Incomplete media''' || '''Supplements for complete media (required)''' | ||
|- | |- | ||
| HEK293 || DMEM high glucose || 10% FBS, 1% pen/strep | | 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 | | K-562 (suspension) || Iscove's Modified Dulbecco's Medium (IMDM) || 10% FBS, 1% pen/strep | ||
| Line 21: | Line 21: | ||
<br> | <br> | ||
==Transgenic | ==Transgenic Cell Selection== | ||
{| class="wikitable" cellspacing=5px | |||
|- valign="top" | |||
| '''Cell Type''' || '''Genotype''' || '''Transfected Plasmid''' || '''Medium''' || '''Supplements<br>(required)''' || '''Selection<br>antibiotic(s)''' | |||
|- | |||
| U2OS (adherent) || Flp-in T-REx || pcDNA5/FRT, or V0200 || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 200 μg/mL hygromycin | |||
|} | |||
<br> | |||
==Transgenic Cell Maintenance== | |||
{| class="wikitable" cellspacing=5px | {| class="wikitable" cellspacing=5px | ||
|- valign="top" | |- valign="top" | ||
| '''Cell Type''' || '''Genotype''' || '''Medium''' || '''Supplements<br>(required)''' || '''Maintenance<br>antibiotic(s)''' | | '''Cell Type''' || '''Genotype''' || '''Medium''' || '''Supplements<br>(required)''' || '''Maintenance<br>antibiotic(s)''' | ||
|- | |- | ||
| U2OS (adherent) || Flp-in T-REx || McCoy's 5A || 10% FBS, 1 % pen/strep || | | U2OS (adherent) || Flp-in T-REx, untransfected || McCoy's 5A || 10% FBS, 1 % pen/strep || 100 ug/mL Zeocin, 15 ug/mL blasticidin | ||
|- | |||
| U2OS (adherent) || Flp-in T-REx, hygro-transfected || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 60 μg/mL hygromycin | |||
|- | |||
| U2OS (adherent) || Flp-in T-REx, puro-transfected || McCoy's 5A || 10% FBS, 1 % pen/strep || 15 ug/mL blasticidin, 0.5 μg/mL puromycin | |||
|- | |||
| HEK 293 (adherent) || Flp-in T-REx || DMEM, high glucose || 10% FBS, 1 % pen/strep || 100 ug/mL Zeocin, 15 ug/mL blasticidin | |||
|- | |- | ||
| HEK 293 (adherent) || | | HEK 293 (adherent) || luc Gal4-EED "23;4;9" (K. Hansen) || DMEM, high glucose || 10% FBS, 1 % pen/strep || 0.5 ug/mL puromycin | ||
|} | |} | ||
Revision as of 19:00, 9 July 2014
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 |
| U2OS (adherent) | McCoy's 5A | 10% FBS, 1% pen/strep |
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, 1 % pen/strep | 15 ug/mL blasticidin, 200 μg/mL hygromycin |
Transgenic Cell Maintenance
| Cell Type | Genotype | Medium | Supplements (required) |
Maintenance antibiotic(s) |
| U2OS (adherent) | Flp-in T-REx, untransfected | McCoy's 5A | 10% FBS, 1 % pen/strep | 100 ug/mL Zeocin, 15 ug/mL blasticidin |
| U2OS (adherent) | Flp-in T-REx, hygro-transfected | McCoy's 5A | 10% FBS, 1 % pen/strep | 15 ug/mL blasticidin, 60 μg/mL hygromycin |
| U2OS (adherent) | Flp-in T-REx, puro-transfected | McCoy's 5A | 10% FBS, 1 % pen/strep | 15 ug/mL blasticidin, 0.5 μg/mL puromycin |
| HEK 293 (adherent) | Flp-in T-REx | DMEM, high glucose | 10% FBS, 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, 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
