Difference between revisions of "Haynes:MamCultureMedia"

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(Transgenic Cell Maintenance)
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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
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<br>
 
<br>
  
==Transgenic Cells==
+
==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 || ## ug/mL puromycin
+
| 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) || Flp-in T-REx || DMEM (high glucose) || 10% FBS, 1 % pen/strep || ## ug/mL puromycin
+
| 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

<- Back to Protocols

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