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年限：embryo, blastocyst
運輸方式：凍存運輸
器官來源：胚胎
細胞形態(tài)：球形
ATCC Number：CRL-1934?
數(shù)量：大量
生長狀態(tài)：貼壁生長
細胞類型：其他細胞類型
品系：129S2/SvPas
是否是腫瘤細胞：0
物種來源：小鼠
規(guī)格：0.1ml Designations: ES-D3 [D3]
Depositors: ?T Doetschman
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:spherical colony


Source: Organ: embryo
Strain: 129S2/SvPas
Cell Type: pluripotent embryonic stem cell;
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications:Undifferentiated cells can be genetically modified by gene targeting techniques.
The cells spontaneously differentiate into embryonic structures in the absence of a feeder layer or conditioned medium.
The clonal embryonic stem cell line ES-D3 was derived from blastocysts of a 129S2/SvPas mouse.
Fibroblast-like feeder layer cells are present in the ampules sent by ATCC .
Age: embryo, blastocyst
Comments:The clonal embryonic stem cell line ES-D3 was derived from blastocysts of a 129S2/SvPas mouse. Undifferentiated cells can be genetically modified by gene targeting techniques.The cells spontaneously differentiate into embryonic structures in the absence of a feeder layer or conditioned medium. They can be maintained in the undifferentiated state by frequent subculture (every 2 to 3 days) on confluent feeder layers (STO cells) irradiated at 12,000 Rads (see ATCC 56-X; irradiated STO cells). These ES-D3 cells are not germline competent. Fibroblast-like feeder layer cells are present in the ampules sent by ATCC .
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: 2-mercaptoethanol to a final concentration of 0.1 mM ; fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0℃
Growth Conditions: To maintain the cells in the undifferentiated state they must be grown on confluent feeder layers of irradiated STO cells (see 56-X, irradiated STO cells).
Subculturing: Protocol:

1. Plate irradiated (12,000 Rads) STO feeder layer at approximately 5.0 to 6.0 X 10(6) cells/ 75 cm2 (confluent monolayer) at least one day before plating the ES cells.
   Note: If the colonies are close to or touching each other the culture is overgrown
2. Remove and discard culture medium.
3. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
4. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37?C to facilitate dispersal.
5. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
6. Add appropriate aliquots of the cell suspension to new culture vessels pre-plated with new feeder layer.
7. Incubate cultures at 37?C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
feeder layer cells:ATCC 56-X
derivative:ATCC SCRC-1003
References: 22394: Doetschman TC, et al. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J. Embryol. Exp. Morphol. 87: 27-45, 1985. PubMed: 3897439
22928: Williams RL, et al. Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature 336: 684-687, 1988. PubMed: 3143916
23296: Gossler A, et al. Transgenesis by means of blastocyst-derived embryonic stem cell lines. Proc. Natl. Acad. Sci. USA 83: 9065-9069, 1986. PubMed: 3024164
23307: Doetschman T, et al. Targeted mutation of the Hprt gene in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 85: 8583-8587, 1988. PubMed: 3186749