07072001
ES-R1 from mouse
07072001, mouse embryo. Monolayer of spheroidal cells on feeder layer of PMEFs.
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About This Item
Código UNSPSC:
41106514
NACRES:
NA.81
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product name
ES-R1 from mouse,
origen biológico
mouse embryo
formulario
liquid
modo de crecimiento
Adherent
cariotipo
Not specified
morfología
Adherent monolayer of spheroidal cells on feeder layer of mouse primary embryonic fibroblasts
productos
Not specified
receptores
Not specified
técnicas
cell culture | mammalian: suitable
Condiciones de envío
dry ice
temp. de almacenamiento
−196°C
Origen línea celular
Mouse embryonic stem cell
Descripción línea celular
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/+).
Medio de cultivo
MEF medium consists of Advanced DMEM/F12 (Invitrogen 12634010), 10% FBS (Perbio SH30070.03E), 2 mM Glutamine (Invitrogen 25030024) and 0.1 mM ß-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 ß-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 ß-mercaptoethanol (Sigma M6250) and LIF 1000 Units/ml (ESGRO ESG1106).
Rutina de subcultivo
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.
Otras notas
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.
Código de clase de almacenamiento
10 - Combustible liquids
Clase de riesgo para el agua (WGK)
WGK 3
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
Certificados de análisis (COA)
Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»
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A Nagy et al.
Proceedings of the National Academy of Sciences of the United States of America, 90(18), 8424-8428 (1993-09-15)
Several newly generated mouse embryonic stem (ES) cell lines were tested for their ability to produce completely ES cell-derived mice at early passage numbers by ES cell <==> tetraploid embryo aggregation. One line, designated R1, produced live offspring which were
Anne Plück et al.
Methods in molecular biology (Clifton, N.J.), 561, 199-217 (2009-06-09)
Since the technique of introducing a targeted mutation ('gene targeting') into the mouse genome was published almost 20 years ago (Cell 51:503-512, 1987), the number of mouse mutants (mouse models) is increasing, especially after the advent of the full mouse
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