MESC 17

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.

• Sinonimo/i: MESC-17, MESC17
• Codice UNSPSC: 41106514

Proprietà

• Origine biologica: mouse embryo
• Confezionamento: tube of 5 μg 11120817-DNA-5UG; pkg of vial of cells 11120817-1VL
• Modalità di accrescimento: Adherent
• Cariotipo: XX, diploid
• Morfologia: Spheroidal
• Prodotti: Not specified
• Recettori: Not specified
• tecniche: cell culture | mammalian: suitable
• Condizioni di spedizione: dry ice

Descrizione

Origine della linea cellulare

Mouse embryonic stem cell

Descrizione della linea cellulare

The germ-line competent cell line MESC 17 was established from the inner cell mass of a 3.5 day female pre-implantation mouse embryo (strain C57BL/6J). These pluripotent cells retain the ability to participate in normal embryonic development.

Profilo DNA

Not specified

Terreno di coltura

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 product number 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 product number M6250) and LIF 1000 Units/ml (ESGRO ESG1106).

Mantenimento delle subcolture

The MESC lines can be grown without the use of mitotically inactivated feeder cells (Brown et al., 1992 PMID: 1483967). However, the cells supplied by ECACC have been grown on mitomycin treated primary mouse embryonic fibroblasts to ensure the cells are maintained in an undifferentiated state. Mouse embryonic fibroblasts, STO (Sigma product number 86032003) or SNL 76/7 (Sigma product number 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/500ml) 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 min 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 h 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 min at room temperature. 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 min. 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.

Altre note

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