ZEM2S細(xì)胞

運輸方式： 凍存運輸

生長狀態(tài)： 貼壁生長

ATCC Number： CRL-2147?

數(shù)量： 大量

細(xì)胞形態(tài)： 成纖維樣

器官來源： 胚胎

是否是腫瘤細(xì)胞： 0

物種來源： 斑馬魚

年限： embryo; blastula embryo, blastula

Designations: ZEM2S

ZEM2S細(xì)胞Depositors: DW Barnes

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Danio rerio deposited as Brachydanio rerio

Morphology: fibroblast

Source: Organ: embryo

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Age: embryo; blastula embryo, blastula

Comments: ZEM2S細(xì)胞ZEM2S was derived from the ZEM2 cell line that had been established from zebrafish embryos using a complex growth medium supplemented with insulin, trout embryo extract, trout and fetal bovine sera.

And medium conditioned by cells from the buffalo rat liver cell line (BRL 3A, see ATCC CRL-1442).

ZEM2S cells were derived from ZEM2 in 1993 by selection for growth in a basal nutrient medium supplemented with 5 to 10% heat-inactivated fetal bovine serum.

These cells provide an in vitro assay system for the comparison of enhancer/promotor activities in early stage zebrafish embryos.

Both transient and stable expression of foreign genes have been demonstrated in transfected zebrafish blastula derived cell cultures.

The cells may provide an in vitro assay system for the study of extracelluar factors which induce and regulate cell differentiation.

A culture submitted to the ATCC in October 1994 was found to be contaminated with mycoplasma, and progeny were cured by a 21 day treatment with BM Cycline.

Propagation: ATCC complete growth medium:

50% Leibovitz's L-15 medium (ATCC 30-2008)

35% Dulbecco's Modified Eagle's medium, high glucose (GIBCO 12100)

15% F12 medium (GIBCO 21700)

The above are all without sodium bicarbonate

Supplemented with:

0.18 g/L sodium bicarbonate

15 mM HEPES

10% heat-inactivated fetal bovine serum

Temperature: 28.0°C

ZEM2S細(xì)胞Max Temperature: 29.0°C

Min Temperature: 26.0°C

Subculturing: Protocol:

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 28?C to facilitate dispersal.

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

To remove Trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.

Discard supernatant and resuspend cells in fresh growth medium.

Add appropriate aliquots of cell suspension to new culture vessels.

Incubate cultures at 28?C without CO2 for 30 minutes.

Examine to ensure attachment, and then add heat-inactivated FBS at 10% of total volume.

Incubate cultures at 28?C without CO2

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended

Medium Renewal: Twice per week

Preservation: Freeze medium: Complete growth medium, 85%; additional HI-FBS, 10%, DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

References: 22330: Collodi P, et al. ZEM2S細(xì)胞Culture of cells from zebrafish (Brachydanio rerio) embryo and ***** tissues. Cell Biol. Toxicol. 8: 43-61, 1992. PubMed: 1591622

22454: Ghosh C, Collodi P. Culture of cells from zebrafish (Brachydanio rerio) blastula-stage embryos. Cytotechnology 14: 21-26, 1994. PubMed: 7765109

22606: . . J. Tissue Culture Methods 16: 99-107, 1994.

22755: . . Mol. Mar. Biol. Biotechnol. 1: 426-431, 1992.