EA.hy926 (ATCC® CRL-2922)

Organism: Homo sapiens, human  /  Cell Type: Endothelial  /  Tissue: somatic cell hybrid  / 

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Organism Homo sapiens, human
Tissue somatic cell hybrid
Cell Type Endothelial
Product Format frozen
Morphology endothelial
Culture Properties adherent
Biosafety Level 1
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
The human umbilical vein cell line, EA.hy926, was established by fusing primary human umbilical vein cells with a thioguanine-resistant clone of A549 by exposure to polyethylene glycol (PEG). Hybrid clones were selected in HAT medium and screened for factor VIII-related antigen.
Antigen Expression
Factor VIII-related antigen; Homo sapiens, expressed
Comments

Electron photomicrographs demonstrate cytoplasmic distribution of Weibel-Palade bodies and tissue-specific organelles, characteristics of differentiated endothelial cell functions such as angiogenesis, homeostasis/thrombosis, blood pressure and inflammation. RefEdgell CJ, et al. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl. Acad. Sci. USA 80: 3734-3737, 1983. PubMed: 6407019 RefEdgell CJ, et al. Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA.hy926. In Vitro Cell. Dev. Biol. 26: 1167-1172, 1990. PubMed: 2079463

EA.hy926 cells have been maintained for more than 100 population doublings (PDLs).

Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. 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 75 cm2 flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 2 X 103 to 3 X 103 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C. Subculture when cell concentration reaches between 8 X 104 and 1 X 105 cells/cm2.

Interval: Twice a week
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended
Medium Renewal: Every 2 to 3 days

Cryopreservation
Freeze medium: Complete Growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 10,11,12
D13S317: 11
D16S539: 11,12
D5S818: 11
D7S820: 8,9,10
THO1: 6,8,9.3
TPOX: 8,9
vWA: 14,17
Name of Depositor CS Edgell
References

Edgell CJ, et al. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl. Acad. Sci. USA 80: 3734-3737, 1983. PubMed: 6407019

Bauer J, et al. In vitro model of angiogenesis using a human endothelium-derived permanent cell line: contributions of induced gene expression, G-proteins, and integrins. J. Cell. Physiol. 153: 437-449, 1992. PubMed: 1280276

Edgell CJ, et al. Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA.hy926. In Vitro Cell. Dev. Biol. 26: 1167-1172, 1990. PubMed: 2079463

Rieber AJ, et al. Extent of differentiated gene expression in the human endothelium-derived EA.hy926 cell line. Thromb. Haemostasis 69: 476-480, 1993. PubMed: 8322270

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

Edgell CJ, et al. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl. Acad. Sci. USA 80: 3734-3737, 1983. PubMed: 6407019

Bauer J, et al. In vitro model of angiogenesis using a human endothelium-derived permanent cell line: contributions of induced gene expression, G-proteins, and integrins. J. Cell. Physiol. 153: 437-449, 1992. PubMed: 1280276

Edgell CJ, et al. Endothelium specific Weibel-Palade bodies in a continuous human cell line, EA.hy926. In Vitro Cell. Dev. Biol. 26: 1167-1172, 1990. PubMed: 2079463

Rieber AJ, et al. Extent of differentiated gene expression in the human endothelium-derived EA.hy926 cell line. Thromb. Haemostasis 69: 476-480, 1993. PubMed: 8322270