ES-D3細(xì)胞

細(xì)胞形態(tài)： 其他

生長狀態(tài)： 貼壁生長

年限： embryo, blastocyst

運輸方式： 凍存運輸

品系： 129S2/SvPas

細(xì)胞類型： 其他細(xì)胞類型

數(shù)量： 大量

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

ATCC Number： CRL-11632?

器官來源： 胚胎

Designations: ES-D3 [D3]

Depositors: P Chambon

ES-D3細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: spherical colonies

Source: Organ: embryo

Strain: 129S2/SvPas

Cell Type: pluripotent embryonic stem cell;

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Age: embryo, blastocyst

Comments: The cells spontaneously differentiate into embryonic structures in the absence of a feeder layer or conditioned medium. They can be injected back into blastocysts and contribute to the germline. Undifferentiated cells can be genetically modified by gene targeting techniques.

Propagation: ES-D3細(xì)胞ATCC complete growth medium: Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose supplemented with 0.1 mM 2-mercaptoethanol, 85%; heat-inactivated fetal bovine serum, 15%

Temperature: 37.0°C

Subculturing: Protocol: Note: The cells can be maintained in the undifferentiated state by frequent subculture on feeder layers of irradiated (3000 rads) or mitomycin C treated (0.01 mg/ml for 90 minutes) primary mouse embryonic fibroblasts or STO cells (see ATCC CRL-1503, STO or ATCC 56-X, irradiated STO cells).

Remove and discard culture medium.

Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.

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.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Add fresh medium, aspirate and dispense onto fresh feeder layer cultures.

Incubate cultures at 37?C.

ES-D3細(xì)胞Subcultivation Ratio: A subcultivation ratio of 1:4 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

feeder layer cells:ATCC 56-X

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. ES-D3細(xì)胞Targeted mutation of the Hprt gene in mouse embryonic stem cells. Proc. Natl. Acad. Sci. USA 85: 8583-8587, 1988. PubMed: 3186749

70349: Chambon P, et al. Genetically engineered mice containing alterations in the genes encoding retinoic acid receptor proteins. US Patent 6,486,381 dated Nov 26 2002