CGR8

NOTE: Both the cell line and DNA from the cell line may be available for this product. Please choose -1VL or VIAL for cells, or -DNA-5UG for DNA.

• UNSPSC Code: 41106514
• Biological source: mouse embryo
• Growth mode: Adherent
• Karyotype: 40XY
• Morphology: Not specified
• Manufacturer/tradename: ECACC

Eigenschaften

• biological source: mouse embryo
• manufacturer/tradename: ECACC
• packaging: tube of 5 μg 07032901-DNA-5UG, pkg of vial of cells 07032901-1VL
• growth mode: Adherent
• karyotype: 40XY
• morphology: Not specified
• products: Not specified
• receptors: Not specified
• technique(s): cell culture | mammalian: suitable
• shipped in: dry ice
• storage temp.: −196°C

Beschreibung

Application

Gene targeting, Gene trapping, in vitro differentiation

Biochem/physiol Actions

Mouse embryonic stem cell

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.

Preparation Note

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

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.

Other Notes

Additional freight & handling charges may be applicable for Asia-Pacific shipments. Please check with your local Customer Service representative for more information.

This cell line is part of the European Collection of Authenticated Cell Cultures (ECACC), an international repository managed by the United Kingdom Health Security Agency (UKHSA). No licensing agreement is required when either this cell line or the DNA extracted from it are used for internal research purposes only. Any other use of these products is prohibited without the express written permission of UKHSA. Inquiries regarding authorized use of this cell line or its genomic DNA may be directed to culturecollections@ukhsa.gov.uk.