產(chǎn)品名稱 |
ZEM2S |
商品貨號 |
B165991 |
Organism |
Danio rerio, zebrafish |
Tissue |
embryo |
Product Format |
frozen |
Morphology |
fibroblast |
Culture Properties |
adherent |
Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
Age |
embryo; blastula |
Applications |
These cells provide an in vitro assay system for the comparison of enhancer/promotor activities in early stage zebrafish embryos.
The cells may provide an in vitro assay system for the study of extracelluar factors which induce and regulate cell differentiation.
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Storage Conditions |
liquid nitrogen vapor phase |
Images |
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Derivation |
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.
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. |
Comments |
Medium conditioned by cells from the buffalo rat liver cell line available as BRL 3A (see ATCC CRL-1442).
Both transient and stable expression of foreign genes have been demonstrated in transfected zebrafish blastula derived cell cultures.
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. |
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
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Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
- Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 mL of SERUM FREE growth medium and aspirate cells by gently pipetting.
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To remove Trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
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Discard supernatant and resuspend cells in fresh growth medium.
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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 |
Cryopreservation |
Freeze medium: Complete growth medium, 85%; additional HI-FBS, 10%, DMSO, 5% Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Temperature: 28°C
Max Temperature: 29°C
Min Temperature: 26°C |
Name of Depositor |
DW Barnes |
Deposited As |
Brachydanio rerio |
Year of Origin |
1993 |
References |
Collodi P, et al. Culture of cells from zebrafish (Brachydanio rerio) embryo and adult tissues. Cell Biol. Toxicol. 8: 43-61, 1992. PubMed: 1591622
Ghosh C, Collodi P. Culture of cells from zebrafish (Brachydanio rerio) blastula-stage embryos. Cytotechnology 14: 21-26, 1994. PubMed: 7765109
Bradford CS, et al. Cell Cultures from zebrafish embryos and adult tissues. J. Tissue Culture Methods 16: 99-107, 1994.
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