CGR8

7032901, mouse embryo, Not specified

• Código UNSPSC: 41106514

Propiedades

• product name: CGR8, 07032901
• origen biológico: mouse embryo
• modo de crecimiento: Adherent
• cariotipo: 40XY
• morfología: Not specified
• productos: Not specified
• receptores: Not specified
• técnicas: cell culture | mammalian: suitable
• Condiciones de envío: dry ice
• temp. de almacenamiento: −196°C

Descripción

Origen línea celular

Mouse embryonic stem cell

Descripción línea celular

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.

Aplicación

Gene targeting, Gene trapping, in vitro differentiation

Medio de cultivo

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

Rutina de subcultivo

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.

Otras notas

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