HUV-EC-C [HUVEC] (ATCC® CRL-1730)

Organism: Homo sapiens, human  /  Cell Type: endothelial  /  Tissue: umbilical vein/vascular endothelium  /  Disease: normal

Organism Homo sapiens, human
Tissue umbilical vein/vascular endothelium
Cell Type endothelial
Product Format frozen
Morphology endothelial
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.

Disease normal
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Karyotype Karyology performed for one batch of CRL-1730 in 1996 reflected a hypodiploid human cell line with a modal chromosome number of 45 occurring in 72% of the cells counted, all of which had monosomic N13. The rate of polyploid cells among this population was 15.8%. This karyology differed from earlier work-ups performed on the cells that showed approximately 60% of the cells retained 2 chromosomes 13. The apparent clonal variation in cultures of CRL-1730 (most likely dependent upon passage and growth conditions) has also been noted in STR profiles with unstable alleles at D13S317 allele #9, D13S317 allele #11, and D7S820 allele #12. Other coexisting subclones include those with 46,XX,-11,-13,i(11p),i(11q) and 46,XX,+11,-13 karyotypes. For all karyotypes performed, both X chromosomes appear normal.
Images
Genes Expressed
factor VIII
Cellular Products
factor VIII
Tumorigenic No
Effects
Yes, the cells did form colonies in semisolid medium.
No, the cells were not tumorigenic in immunosuppressed mice
Comments
Endothelial Cell Growth Supplement (ECGS) and unidentified factors from bovine pituitary, hypothalamus or whole brain extracts are mitogenic for this line.
The cells have a life expectancy of 50 to 60 population doublings.
Complete Growth Medium

The base medium for this cell line is F-12K Medium (ATCC 30-2004). To make the complete growth medium, add the following components to 440 mL of the base medium:

  • 5 mL of a 10 mg/mL stock heparin solution (prepared from Sigma catalog #H3393) for a final concentration of 0.1 mg/mL heparin in complete growth medium 
    • Dissolve 1 g Heparin in 100 mL basal F-12K  and filter to make a 10 mg/mL stock solution
  • 5 mL endothelial cell growth supplement (ECGS; BD Biosciences catalog # 354006)
    • Rehydrate 1 vial using 5 mL basal F-12K 
  • 50 mL fetal bovine serum (FBS; ATCC 30-2020)
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. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.

Note: A high quality ECGS prepared from bovine neural tissue (BD Biosciences catalog # 354006 or equivalent) should be used to propagate CRL-1730. It is best to initiate the cells with the highest recommended concentration of ECGS. Moderate to heavy debris and numerous floating cells may be routinely observed in cultures of HUV-EC-C cells.  Retain the floating cells by gentle centrifugation and add back to the adherent population.

CRL-1730 is a slow growing cell line that has a roughly estimated doubling time of 5 to 6 days. Cultures should be fully fluid changed every 48 hours. The cells should only be allowed to go 72 hours without fluid changing when the density is less than 50% confluent. Perform full fluid changes rather than media additions.

This cell line produces a lot of floaters and debris especially at higher densities. Cells detach before completely filling in to 100% confluence. It is recommended to subculture the cells when 80 to 90% confluent to avoid excessive floaters. Floating cells are viable and if pronounced, they should be spun down and reseeded back into the growing culture.

  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 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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

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

Medium Renewal: Two to three times per week

Seeding Density: 8.0 x 103 to 3.0 x 104 viable cells/cm2

Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 11,12
D13S317: 9,11
D16S539: 11,12
D5S818: 11,12
D7S820: 8,12
THO1: 6,9.3
TPOX: 8,11
vWA: 16
Name of Depositor H Hoshi
References

Molestina RE, et al. Characterization of a strain of Chlamydia pneumoniae isolated from a coronary atheroma by analysis of the omp1 gene and biological activity in human endothelial cells. Infect. Immun. 66: 1370-1376, 1998. PubMed: 9529055

Zahedi K. Characterization of the binding of serum amyloid P to laminin. J. Biol. Chem. 272: 2143-2148, 1997. PubMed: 8999915

Lindstrom AL, et al. An in vitro model for toxin-mediated vascular leak syndrome: ricin toxin A chain increases the permeability of human endothelial cell monolayers. Blood 90: 2323-2334, 1997. PubMed: 9310483

Soker S, et al. Inhibition of vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation by a peptide corresponding to the exon 7-encoded domain VGEF165. J. Biol. Chem. 272: 31582-31588, 1997. PubMed: 9395496

Li Y, et al. Mast cell granules potentiate endotoxin-induced interleukin-6 production by endothelial cells. J. Leukocyte Biol. 62: 211-216, 1997. PubMed: 9261335

Soker S, et al. Characterization of novel vascular endothelial growth factor (VEGF) receptors on tumor cells that bind VEGF165 via its exon 7-endoded domain. J. Biol. Chem. 271: 5761-5767, 1996. PubMed: 8621443

Hoshi H, McKeehan WL. Brain- and liver cell-derived factors are required for growth of human endothelial cells in serum-free culture. Proc. Natl. Acad. Sci. USA 81: 6413-6417, 1984. PubMed: 6333682

Basic Documentation
Other Documentation
FAQ's
  1. ATCC® CRL-1730 vs. ATCC® CRL-2873


    Date Updated: 2/4/2013

  2. Number of ATCC® CRL-1730 cells in a confluent flask


    Date Updated: 2/5/2013

  3. CRL-1730 morphology and culture condition


    Date Updated: 2/21/2014

  4. ATCC HUVEC lines
    Human umbilical vein endothelial cells, termed HUVEC (ATCC® PCS-100-010 ™), are primary cells that are distributed by ATCC as part of the Primary Cell Solutions™ cell offerin...
    Date Updated: 3/27/2014
  5. Tech Q Challenge (ATCC Connection, 30(2):19)

    EC = Endothelial Cells


    Date Updated: 7/25/2012
References

Molestina RE, et al. Characterization of a strain of Chlamydia pneumoniae isolated from a coronary atheroma by analysis of the omp1 gene and biological activity in human endothelial cells. Infect. Immun. 66: 1370-1376, 1998. PubMed: 9529055

Zahedi K. Characterization of the binding of serum amyloid P to laminin. J. Biol. Chem. 272: 2143-2148, 1997. PubMed: 8999915

Lindstrom AL, et al. An in vitro model for toxin-mediated vascular leak syndrome: ricin toxin A chain increases the permeability of human endothelial cell monolayers. Blood 90: 2323-2334, 1997. PubMed: 9310483

Soker S, et al. Inhibition of vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation by a peptide corresponding to the exon 7-encoded domain VGEF165. J. Biol. Chem. 272: 31582-31588, 1997. PubMed: 9395496

Li Y, et al. Mast cell granules potentiate endotoxin-induced interleukin-6 production by endothelial cells. J. Leukocyte Biol. 62: 211-216, 1997. PubMed: 9261335

Soker S, et al. Characterization of novel vascular endothelial growth factor (VEGF) receptors on tumor cells that bind VEGF165 via its exon 7-endoded domain. J. Biol. Chem. 271: 5761-5767, 1996. PubMed: 8621443

Hoshi H, McKeehan WL. Brain- and liver cell-derived factors are required for growth of human endothelial cells in serum-free culture. Proc. Natl. Acad. Sci. USA 81: 6413-6417, 1984. PubMed: 6333682