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UK Health Security Agency

CGR8

CGR8

Catalogue No.

07032901

Cell Line Name

CGR8

Cell Line Description

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 is essential and allows culture of CGR8 without the use of feeder layers. Cells are small and tightly packed.

Characteristics

Tissue of Origin

Embryo

Karyotype

40XY

Applications

Gene targeting, Gene trapping, in vitro differentiation

Disease

None Stated

Culture Conditions

Subculture Routine

Split sub-confluent cultures (70-80%) 1:10 i.e. seeding at 4x1000 cells/cm² using 0.05% 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.

Culture Medium

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

Growth Mode

Adherent

Additional Info

Depositor

Dr A Smith, AFRC Centre for Genome Research, Edinburgh

Country of Origin

United Kingdom

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

J Tiss Cult Meth 1991;13:89; Proc Natl Acad Sci USA 1994;4303; Development 1990;110:1341

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: CGR8 (ECACC 07032901).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.