Haynes:MamCultureMedia: Difference between revisions
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<- [[Haynes:Protocols | Back to Protocols]] | |||
<div style="width: 1000px"> | |||
=Mammalian Culture Media= | =Mammalian Culture Media= | ||
=="Wild type" stock cells (non-transgenic)== | =="Wild type" stock cells (non-transgenic)== | ||
{| class="wikitable" | {| class="wikitable" cellspacing=5px | ||
|- | |- | ||
| '''Cell line''' || '''Incomplete media''' || '''Supplements for complete media (required)''' | | '''Cell line''' || '''Incomplete media''' || '''Supplements for complete media (required)''' | ||
|- | |- | ||
| U2OS (adherent) || McCoy's 5A || 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 | |||
|- | |||
| 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 | |||
|} | |||
<br> | |||
==Transgenic Cells== | |||
{| class="wikitable" cellspacing=5px | |||
|- valign="top" | |||
| '''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 || 100 ug/mL Zeocin, 15 ug/mL blasticidin | |||
|- | |- | ||
| | | 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 | |||
|} | |} | ||
<br> | |||
==Notes== | |||
* FBS = fetal bovine serum | * FBS = fetal bovine serum | ||
| Line 18: | Line 43: | ||
* Store all media at 4°C | * Store all media at 4°C | ||
</div> | |||
Revision as of 19:16, 22 February 2013
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 Cells
| Cell Type | Genotype | Medium | Supplements (required) |
Maintenance antibiotic(s) |
| U2OS (adherent) | Flp-in T-REx | McCoy's 5A | 10% FBS, 1 % pen/strep | 100 ug/mL Zeocin, 15 ug/mL blasticidin |
| 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
