運輸方式：凍存運輸
ATCC Number：CRL-1730?
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生長狀態(tài)：貼壁生長
數(shù)量：大量
組織來源：vascular endothelium
細胞形態(tài)：內皮樣
器官來源：其他
細胞類型：內皮細胞
是否是腫瘤細胞：0
物種來源：人
規(guī)格：0.1ml Designations: HUV-EC-C
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens
Morphology:endothelial
Source: Organ: umbilical vein
Tissue: vascular endothelium
Disease: normal
Cell Type: endothelial
Cellular Products:factor VIII [23284 ]
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Applications:transfection host
Tumorigenic:No
DNA Profile (STR):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
Cytogenetic Analysis: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.
Comments:Endothelial Cell Growth Supplement (ECGS) and unidentified factors from bovine pituitary, hypothalamus or whole brain extracts are mitogenic for this line. [23284 ]
The cells have a life expectancy of 50 to 60 population doublings.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated of F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: 0.1 mg/ml heparin; 0.03-0.05 mg/ml endothelial cell growth supplement (ECGS); adjust to a final concentration of 10% fetal bovine serum.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Note: A high quality ECGS prepared from bovine neural tissue (Sigma Cat no. E-2759 or equivalent) should be used to propagate CRL-1730. It is best to initiate the cells with the highest recommended concentration of ECGS.Protocol:
- Remove and discard culture medium.
- 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.
- 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. - Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- 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
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2004
recommended serum:ATCC 30-2020
References: 23284: 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
29192: Zahedi K. Characterization of the binding of serum amyloid P to laminin. J. Biol. Chem. 272: 2143-2148, 1997. PubMed: 8999915
33021: 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
