CGR8

7032901, mouse embryo, Not specified

• Code UNSPSC : 41106514

Propriétés

• product name: CGR8, 07032901
• Source biologique: mouse embryo
• Mode de croissance: Adherent
• Caryotype: 40XY
• Morphologie: Not specified
• Produits: Not specified
• Récepteurs: Not specified
• Technique(s): cell culture | mammalian: suitable
• Conditions d'expédition: dry ice
• Température de stockage: −196°C

Description

Origine de la lignée cellulaire

Mouse embryonic stem cell

Description de la lignée cellulaire

The germ-line competent cell line CGR8 was established from the inner cell mass of a 3.5 day male pre-implantation mouse embryo (Mus musculus, strain 129). These pluripotent cells retain the ability to participate in normal embryonic development. Differentiation of CGR8 cells is inhibited by the pleiotropic cytokine Differentiation Inhibiting Activity (DIA) which is identical to Leukaemia Inhibiting Factor (LIF). Addition of DIA/LIF allows culture of CGR8 without the use of feeder layers. Cells are small and tightly packed.

Application

Gene targeting, Gene trapping, in vitro differentiation

Milieu de culture

GMEM + 2mM Glutamine + 0.05mM 2-Mercaptoethanol (2ME) + 1000 units/ml DIA/LIF + 10% Foetal Bovine Serum (FBS).

Procédure de repiquage

Split sub-confluent cultures (70-80%) 1:10 i.e. seeding at 4x1000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO2; 37°C. CGR8 cells should be cultured on gelatin - coated flasks. Flasks should be coated using 0.2% gelatin in PBS. ECACC introduced the use of a feeder layer of mitomycin C treated primary mouse embryonic fibroblast (PMEF) cells for the bulk culture of CGR8. This means the ampoules that we provide contain the PMEF feeder cells as well as the CGR8 cells. When the cells are initially resuscitated and plated out from the ampoule both the CGR8 stem cell colonies and the fibroblast feeder cells will be visible in the culture. Feeder layers are not essential and the use of LIF at the correct concentration (with daily media changes) should be sufficient to maintain the pluripotency of the cells in end user laboratories.

Autres remarques

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