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CGR8

7032901, mouse embryo, Not specified

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About This Item

UNSPSCコード:
41106514

製品名

CGR8, 07032901

由来生物

mouse embryo

包装

tube of 5 μg 07032901-DNA-5UG
pkg of vial of cells 07032901-1VL

成長モード

Adherent

核型

40XY

形態

Not specified

製品

Not specified

受容体

Not specified

テクニック

cell culture | mammalian: suitable

輸送温度

dry ice

保管温度

−196°C

細胞株の由来

Mouse embryonic stem cell

細胞株の説明

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.

アプリケーション

Gene targeting, Gene trapping, in vitro differentiation

培地

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

継代と培養方法

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.

その他情報

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