07072001

ES-R1 from mouse

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, 07072001, mouse embryo. Monolayer of spheroidal cells on feeder layer of PMEFs.

• NACRES: NA.81
• UNSPSC Code: 41106514
• Biological source: mouse embryo
• Growth mode: Adherent
• Karyotype: Not specified
• Morphology: Adherent monolayer of spheroidal cells on feeder layer of mouse primary embryonic fibroblasts

Eigenschaften

• Quality Segment: 100
• biological source: mouse embryo
• form: liquid
• packaging: tube of 5 μg 07072001-DNA-5UG, pkg of vial of cells 07072001-1VL
• growth mode: Adherent
• karyotype: Not specified
• morphology: Adherent monolayer of spheroidal cells on feeder layer of mouse primary embryonic fibroblasts
• products: Not specified
• receptors: Not specified
• technique(s): cell culture | mammalian: suitable
• shipped in: dry ice
• storage temp.: −196°C

Beschreibung

Biochem/physiol Actions

Mouse embryonic stem cell

Pluripotent mouse embryonic stem cell line. R1 ES cells were established in August 1991 from a male blastocyst hybrid of two 129 substrains (129X1/SvJ and 129S1/SV-+^p+^Tyr-c Kitl ^Sl-J/+).

Packaging

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.

Preparation Note

Embryonic stem (ES) cells require the use of mitotically inactivated feeder cells to support the growth of stem cells in the undifferentiated state. Mouse embryonic fibroblasts, STO (Sigma catalog no. 86032003) or SNL 76/7 (Sigma catalog no. 07032801) can be used. At ECACC plastic ware is pre-coated with gelatine prior to plating feeder cells.

Porcine gelatine (Sigma G1890) is dissolved in sterile water (0.5 g/500 ml) at 56 °C. The 0.1% solution is sterilized by filtration (0.22 μm). Add 0.1% gelatine to plastic ware to cover bottom, and incubate for 20 minutes at room temperature. Remove gelatine, wash with PBS once and replace with appropriate culture medium. The flask/dish must not be allowed to dry out.

Feeder layers are prepared on the gelatinized flasks at least 24 hours in advance of being required. An ampoule is thawed in 37 °C water bath and the contents quickly transferred to a 15 ml centrifuge tube. MEF medium is added drop wise to 5 ml. Cells are centrifuged at 150 x g for 5 minutes at room temperature (RT). Cells are resuspended in 5 ml of MEF medium. Cells are counted and added to flasks containing the correct medium at 1-3 x 10⁴ cells/cm².

An ampoule of ES cells is thawed in 37 °C water bath and the contents quickly transferred to a 15 ml centrifuge tube. KSR medium is added drop wise to 5 ml. Cells are centrifuged at 150 x g for 5 minutes. Cells are resuspended in 5 ml of KSR medium. The prepared feeder flask is washed once with PBS and KSR medium added. ES cells should be plated at 4-5 x 10⁴ cells/cm². Cultures must be incubated in a humidified 5% CO₂/95% air incubator at 37 °C. A 100% media change must be performed every day and cells passaged every 2-3 days. Colonies must not be allowed to touch each other as overgrowth will result in differentiation.

MEF medium consists of Advanced DMEM/F12 (Invitrogen 12634010), 10% FBS (Perbio SH30070.03E), 2 mM Glutamine (Invitrogen 25030024) and 0.1 mM β-mercaptoethanol (Sigma M6250).

KSR medium consists of KO-DMEM (Gibco 10829), 20% Knock-Out Serum Replacer (Gibco 10828), 2 mM Glutamine (Invitrogen 25030024), NEAA (Invitrogen 11140035), 0.1 mM β-mercaptoethanol (Sigma M6250) and LIF 1000 Units/ml (ESGRO ESG1106).

Other Notes

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

Cultures from HPA Culture Collections and supplied by Sigma 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.

Depositor and originator: Dr A Nagy, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Ave, Toronto, Ontario, M5G 1X5, Canada.

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.

Sicherheitshinweise

• Lagerklasse: 10 - Combustible liquids
• Was ist los?: WGK 3
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable