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運(yùn)輸方式：凍存運(yùn)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
規(guī)格：10ml Designations: ZEM2S
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
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.
Age: embryo; blastula embryo, blastula
Comments: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
Max Temperature: 29.0°C
Min Temperature: 26.0°C
Subculturing: Protocol:

1. Remove and discard culture medium.
2. 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.
3. 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.
4. Add 6.0 to 8.0 ml of SERUM FREE growth medium and aspirate cells by gently pipetting.
5. To remove Trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.
6. Discard supernatant and resuspend cells in fresh growth medium.
7. Add appropriate aliquots of cell suspension to new culture vessels.
8. Incubate cultures at 28?C without CO2 for 30 minutes.
9. Examine to ensure attachment, and then add heat-inactivated FBS at 10% of total volume.
10. 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. Culture of cells from zebrafish (Brachydanio rerio) embryo and adult 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.