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科研细胞
药靶细胞
标准品
HUVEC（人脐静脉上皮细胞）
| CBP60340 | |
| I. General information | |
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Synonyms: |
HUV-EC-C, HUVEC-C, HUVEC |
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Background: |
Normal endothelial cells with limited life span. For cultivation, endothelial cell growth supplement can be substituted by ca 10 ng/ml basic FGF. |
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Species: |
Homo sapiens, human |
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Tissue: |
Umbilical vein/vascular endothelium |
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Disease: |
Normal |
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Gender: |
NA |
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Morphology: |
Epithelial |
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Growth Mode: |
Adherent |
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Doubling Time: |
~48hrs |
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DNA Profile: |
Amelogenin: X Cobioer’s Cell Line Authentication Service |
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Culture Medium: |
BEGM kit+10%FBS |
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Cryopreservation medium: |
90% Complete Medium+10%DMSO |
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Photo: |
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Genes Expressed: |
Factor VIII |
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Effects: |
Yes, the cells did form colonies in semisolid medium. No, the cells were not tumorigenic in immunosuppressed mice. |
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Comments: |
Endothelial Cell Growth Supplement (ECGS) and unidentified factors from bovine pituitary, hypothalamus or whole brain extracts are mitogenic for this line. For more information, please contact Cobioer (4008-750-250). |
II. Handling Procedure for Flask Cultures
| The flask was seeded with cells grown and completely filled with medium at Cobioer to prevent loss of cells during shipping. |
| 1.Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination. |
| Using an inverted microscope (preferably equipped with phasecontrast optics), carefully check for any evidence of microbial contamination. Also check to determine if the majority of cells are still attached to the bottom of the flask; during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable). |
| 2.If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium. The shipping medium can be saved for reuse. Incubate the cells at 37°C in a 5% CO2 in air atmosphere until they are ready to be subcultured. |
| 3.If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 1000rpm for 5 to 10 minutes. Remove shipping medium and save. Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask. Incubate at 37°C in a 5% CO2 in air atmosphere until cells are ready to be subcultured. |
III. Subculturing Procedure
| Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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 that 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 (depend on the batch of trypsin). |
| 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: 2 to 3 times per week |
IV. Cryopreservation Procedure
| Volumes used in this protocol are for 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) Trypsin0.53mM 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 1 to 10 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.To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 1000rpmfor 5 to10 minutes. |
| 6.Discard supernatant and resuspend cells in cryopreservation medium. |
| 7.Transfer the cells into Cryogenic Vials, 1ml/vial. |
| 8.Frozen the cells in cryogenic container (Nalgene #5100-0001). |
V. Database
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Mutation: |
http://www.cobioer.com/sjk/&pmcId=77.html |
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Expression: |
NA |
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Comments: |
For more information, please contact Cobioer (4008-750-250). |
VI. References
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1.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 |
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2.Zahedi K. Characterization of the binding of serum amyloid P to laminin. J. Biol. Chem. 272: 2143-2148, 1997. PubMed: 8999915 |
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3.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 |
| 4.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 |
| 5.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 |
| 6.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 |
| 7.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 |
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