產(chǎn)品名稱 |
EJG |
商品貨號 |
B164401 |
Organism |
Bos taurus, cow |
Tissue |
adrenal gland |
Cell Type |
endothelial |
Product Format |
frozen |
Morphology |
endothelial |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells contain Bovine Viral Diarrhea Virus (BVDV)]
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. |
Disease |
normal |
Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
Derivation |
The line was derived from collagenase dissociated bovine adrenal tissue. |
Genes Expressed |
Factor VIII:C |
Cellular Products |
Factor VIII:C |
Comments |
The cellular morphology is that of capillary endothelium. The cells form linearly oriented confluent cultures. Recent tests for bovine viral diarrhea virus (BVDV) indicate that the line is producing both detectable BVDV antigens and infectious BVDV virions (J. Virol Methods 48:211-221, 1994). |
Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
|
Subculturing |
Volumes used in this protocol are for a 75 cm2 flask? proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of TrypsinEDTA 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.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension (75 sq. cm. with 1 X 104 cells) to new culture vessels.
6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
|
Culture Conditions |
Temperature: 37°C |
Population Doubling Time |
110 hrs |
Name of Depositor |
The United States of America |
U.S. Patent Number |
|
References |
Pollard HB, et al. Isolation and culture of adrenal medullary endothelial cells producing blood clotting factor VIII:C. US Patent 4,670,394 dated Jun 2 1987
Bolin SR, et al. Survey of cell lines in the American Type Culture Collection for bovine viral diarrhea virus. J. Virol. Methods 48: 211-221, 1994. PubMed: 7989438
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