ES-R1-EGFP B5/EGFP
07072007, mouse embryo. Monolayer of spheroidal cells on feeder layer of PMEFs.
Synonym(e):
B5/EGFP, ES-R 1, ES-R-1, ESR-1, ESR1
About This Item
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ES-R1-EGFP B5/EGFP, 07072007, from mouse embryo
Biologische Quelle
mouse embryo
Wachstumsmodus
Adherent
Karyotyp
Not specified
Morphologie
Adherent monolayer of spheroidal cells on feeder layer of mouse primary embryonic fibroblasts
Produkte
Not specified
Rezeptoren
Not specified
Methode(n)
cell culture | mammalian: suitable
Versandbedingung
dry ice
Ursprung der Zelllinie
Beschreibung der Zelllinie
DNA-Profil
Nährmedium
KSR medium consists of KO-DMEM (Gibco 10829), 20% Knock-Out Serum Replacer (Gibco 10828), 2 mM Glutamine (Invitrogen 25030024), NEAA (Invitrogen 11140035), 0.1 mM ß-mercaptoethanol (Sigma M6250) and LIF 1000 Units/ml (ESGRO ESG1106).
Subkultur-Routine
Feeder layers are prepared on the gelatinized flasks at least 24 hours in advance of being required. An ampoule is thawed in 37?C water bath and the contents quickly transferred to a 15ml centrifuge tube. MEF medium is added drop wise to 5ml. Cells are centrifuged at 150 x g for 5 minutes at Room Temperature (RT). Cells are resuspended in 5ml of MEF medium. Cells are counted and added to flasks containing the correct medium at 1-3 x 104 cells/cm2.
An ampoule of ES cells is thawed in 37?C water bath and the contents quickly transferred to a 15ml centrifuge tube. KSR medium is added drop wise to 5ml. Cells are centrifuged at 150 x g for 5 minutes. Cells are resuspended in 5ml of KSR medium. The prepared feeder flask is washed once with PBS and KSR medium added. ES cells should be plated at 4-5 x 104 cells/cm2. Cultures must be incubated in a humidified 5% CO2/95% air incubator at 37?C. A 100% media change must be performed every day and cells passaged every 2-3 days. Colonies must not be allowed to touch each other as overgrowth will result in differentiation.
Sonstige Hinweise
Haftungsausschluss
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