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.
Sign In to View Organizational & Contract Pricing.
Select a Size
Change View
| Size/SKU | Availability | Price |
|---|
About This Item
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
Pricing and availability is not currently available.
Quality Segment
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
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.
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).
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.
1 of 1
This Item | |||
|---|---|---|---|
| description 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. | description frozen liquid (from clotted whole blood) | description recombinant, expressed in NSO cells, ≥90% (SDS-PAGE), lyophilized powder, suitable for cell culture | description recombinant, expressed in E. coli, ≥98% (SDS-PAGE), ≥98% (HPLC), suitable for cell culture |
| growth mode Adherent | growth mode - | growth mode - | growth mode - |
| karyotype Not specified | karyotype - | karyotype - | karyotype - |
| technique(s) cell culture | mammalian: suitable | technique(s) - | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
| biological source mouse embryo | biological source mouse | biological source mouse | biological source mouse |
| receptors Not specified | receptors - | receptors - | receptors - |
| morphology Adherent monolayer of spheroidal cells on feeder layer of mouse primary embryonic fibroblasts | morphology - | morphology - | morphology - |
Storage Class
10 - Combustible liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
Choose from one of the most recent versions:
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.