In Vitro Technologies

Permits and Restrictions	View Permits View Restrictions
Organism	Homo sapiens, human
Tissue	Umbilical vascular endothelium
Cell Type	Endothelial cell immortalized with hTERT
Product Format	frozen 1 mL
Morphology	Cobblestone appearance with large dark nuclei
Culture Properties	adherent
Biosafety Level	2 [These primary cells are not known to harbor an agent recognized to cause disease in healthy adult humans. Handle as a potentially biohazardous material under at least Biosafety Level 1 containment. Cells derived from primate lymphoid tissue may fall under the regulations of 29 CFR 1910.1030 Bloodborne Pathogens. ATCC recommends that appropriate safety procedures be used when handling all primary cells and cell lines, especially those derived from human or other primate material. Detailed discussions of laboratory safety procedures are provided in Laboratory Safety: Principles and Practice, 2nd ed. (ASM Press, Washington, DC) (Fleming et al., 1995) and Caputo, J.L. Biosafety procedures in cell culture. (1988) J. Tissue Culture Methods 11:223. Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.] Human Material Precaution All tissues used for isolation are obtained under informed consent and conform to HIPAA standards to protect the privacy of the donor’s personal health information. It is best to use caution when handling any human cells. We recommend that all human cells be accorded the same level of biosafety consideration as cells known to carry HIV. With infectious virus assays or viral antigen assays, even a negative test result may leave open the possible existence of a latent viral genome. 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
Age	Newborn
Gender	female
Ethnicity	Caucasian
Applications	Immune response, wound healing, cellular response to viral or bacterial infection, oxidative stress, angiogenesis, arteriolosclerosis, drug screening, tubule formation
Shipping Information	Frozen 1 mL
Storage Conditions	-130°C

Karyotype	46,XX,add(14)(p10)
Images
Derivation	Umbilical vascular endothelium transduced with hTERT
Antigen Expression	CD31 >75% positive
Receptor Expression	LDL uptake >75%positive

Complete Growth Medium	The base medium for this cell line is Vascular Cell Basal Medium (ATCC^®PCS-100-030), supplemented with Vascular Endothelial Cell Growth Kit-VEGF (ATCC^®PCS-100-041). Alternatively, the Vascular Cell Basal Medium can be supplemented with Endothelial Cell Growth Kit-BBE (ATCC^®PCS-100-040).
Subculturing	Every 3-4 days at 8000 cells/cm² Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation solutions for culture vessels of other sizes. Remove and discard spent medium. Briefly rinse with 1X PBS (ATCC 30-2200), 1 mL/25 cm2 and discard rinse solution. Add trypsin for primary cells (ATCC PCS-999-003), 1 mL/25 cm². Place at 37^oC for 3-4 minutes, until 90% of the cells have detached. Rapflask gently to ensure cells are detached. Add 2% FBS in D-PBS, 1 mL/25 cm² to neutralize trypsin. Centrifuge cells at 150 x g for 5 min at room temperature. Remove supernatant. Resuspend pellet in 5.0 to 8.0 mL Complete Growth Medium. Count cells, and seed 8.0 x 10³ to 10.0 x 10³ viable cells/cm² to new culture vessels. Medium Renewal: Every 2-3 days.

Complete Growth Medium

The base medium for this cell line is Vascular Cell Basal Medium (ATCC^®PCS-100-030), supplemented with Vascular Endothelial Cell Growth Kit-VEGF (ATCC^®PCS-100-041). Alternatively, the Vascular Cell Basal Medium can be supplemented with Endothelial Cell Growth Kit-BBE (ATCC^®PCS-100-040).

Subculturing

Every 3-4 days at 8000 cells/cm² Volumes used in this protocol are for 75 cm² flasks; proportionally reduce or increase amount of dissociation solutions for culture vessels of other sizes.

Remove and discard spent medium.
Briefly rinse with 1X PBS (ATCC 30-2200), 1 mL/25 cm2 and discard rinse solution.
Add trypsin for primary cells (ATCC PCS-999-003), 1 mL/25 cm². Place at 37^oC for 3-4 minutes, until 90% of the cells have detached.
Rapflask gently to ensure cells are detached. Add 2% FBS in D-PBS, 1 mL/25 cm² to neutralize trypsin.
Centrifuge cells at 150 x g for 5 min at room temperature.
Remove supernatant. Resuspend pellet in 5.0 to 8.0 mL Complete Growth Medium.
Count cells, and seed 8.0 x 10³ to 10.0 x 10³ viable cells/cm² to new culture vessels.

Medium Renewal: Every 2-3 days.

Cells per Vial	1.2 x 10⁶ cells/vial
Volume	1 mL
STR Profile	Amelogenin: X CSF1PO: 12 D5S818: 15, 16 D7S820: 9, 11 D13S317: 11, 12 D16S539: 9, 13 TH01: 6, 9 TPOX: 8, 9 vWA: 16, 19 CO1 Species: Human
Sterility Tests	Bacteria and Yeast: NegativeMycoplasma: Negative
Viability	>70% when thawed from cryopreservation
Population Doubling Capacity	At least 25 PDL
Population Doubling Time	2.5 days

Name of Depositor	Evercyte
Year of Origin	2011

E: care@invitro.com.au
P: 1300 552 003

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HUVEC/TERT 2; Human Umbilical Vein Endothelial Cell; Human (Homo sapiens) Item ID: ATCCRL4053

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