Immortalized Porcine Aortic Endothelial Cells猪主动脉内皮细胞系永生化细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

Immortalized Porcine Aortic Endothelial Cells猪主动脉内皮细胞系永生化细胞株

Immortalized Porcine Aortic Endothelial Cells猪主动脉内皮细胞系永生化细胞株
Cat No.: NTCC-0T0048

Recently proposed as a potential model of atherosclerosis, allograft rejection and accommodation, xenotransplantation and viral haemorrhage diseases, the porcine endothelial cells provide a useful tool in studies including the molecular mechanisms of xenogeneic rejection in the swine-to-human combination, and the pathogenesis of African Swine Fever (ASF) and Classical Swine Fever (CSF). The Immortalized Porcine Aortic Endothelial Cells (AOC) is derived from stable transformation of SV40 genome into primary porcine aortic endothelial cells. The resulting AOC retains comparable surface markers to its primary cell counterparts, as well as preserving its ability to uptake acetylated low density lipoproteins (Ac-LDL) and its susceptibility to xenogeneic cell-mediated cytotoxicity (i.e. Human Natural Killer Cell-mediated lysis).

Cell Image


Cat. No. NTCC®-0T0448
Name Immortalized Porcine Aortic Endothelial Cells (AOC)
Category Immortalized Cell Lines
Cell Type Immortalized Cells
Organism Pig (Porcine)
Tissue Blood Vessel
Cell Morphology Cobble-stone
Growth Properties Adherent
Seeding Density 25,000-30,000 cells/cm2; Recommended split ratio is 1:2 to 1:3
Population Doubling Time 50-60 hours
Immortalization Method Tranfection with pRNS-1 plasmid encoding the SV40 genome
Expression Profile
CD29, CD31, CD41/61, CD80/86, CD46, SWC3, LAMP-1

Propagation Method
Use of PriCoat T25 Flasks (NTCC®-G299) or Applied Cell Extracellular Matrix (NTCC®-G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.
The base medium for this cell line is Prigrow II medium available at NTCC® (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (NTCC®-TM999)* to a final concentration of 20% and Penicillin/Streptomycin Solution (NTCC®-G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0

Subculture Protocol
1. Subculture when the culture reaches 80-85% confluency or above.
2. Warm complete medium, Trypsin-EDTA solution, and DPBS (Ca++ and Mg++ free) to room temperature.
3. Rinse the cells with DPBS.
4. Add 2-3 ml of Trypsin-EDTA solution into flask. Gently rock the flask to ensure complete coverage of cells by T/E solution. Incubate the flask in a 37°C incubator for 1 to 2 minutes or until cells completely round up. Use a microscope to monitor the change in cell morphology.
5. If needed, you may use cell scraper to scrape the cells off from the flask. Caution: make sure to only scrap the cells in one direction motion, avoid scrapping the cells back and forth with the cells scraper blade.
6. Add 6-8 ml of complete medium to the flask and transfer detached cells to the 15 ml centrifuge tube. Rinse the flask with another 3 ml of complete medium to collect the residual cells.
7. Examine the flask under a microscope for a successful cell harvest by looking at the number of cells being left behind; there should be less than 5%.
8. Centrifuge the 15 ml centrifuge tube at 200x g for 2-3 minutes. Resuspend cells in fresh complete culture medium.
9. Count and plate cells in a new prepared culture vessel.

QC
1)Flow cytometry was used to confirm the expression of endothelial cell markers such as CD29, CD31, CD41/61, CD80/86, CD46, SWC3, LAMP-1 antigen and MHC Class I antigens; 2) 51Cr release assay was used to analyse the susceptibility of AOC to xenogeneic cell-mediated cytotoxicity; 3) 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine (DiI)-labelled Ac-LDL was used to evaluate Ac-LDL update by the AOC.

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