Description
Product Description
The pulmonary vasculature is of great physiological/pathological significance. Human Pulmonary Artery Endothelial Cells (HPAEC) are critical to maintain the function of the pulmonary circulation and effective diffusion capacity of the lung. HPAEC provide a useful tool for studying various aspects of pathology and biology of the lung vasculature in vitro [1].
iXCells Biotechnologies provides high quality HPAEC, which are isolated from human pulmonary artery from single donors and cryopreserved at P2, with >0.5 million cells in each vial. HPAEC express vWF/Factor VIII and CD31 (Figure 1) and are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. They can further expand in Endothelial Cell Growth Medium (Cat# MD-0010) under the condition suggested by iXCells Biotechnologies.
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Figure 1. HPAEC provided by iXCells are positive for CD31 (A) and vWF (B), as shown by immunostaining. Flow analysis showed that more than 99% of cells are CD31 positive (A’) and more than 97% are vWF positive (B’).
Product Details
| Tissue | Human pulmonary artery |
| Package Size | 0.5 million cells/vial |
| Passage Number | P2 |
| Shipped | Cryopreserved |
| Storage | Liquid nitrogen |
| Growth Properties | Adherent |
| Media | Endothelial Cell Medium Kit (Cat# MD-0010) |
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Protocols |
| Thawing of Frozen Cells
1. Upon receipt of the frozen cells, it is recommended to thaw the cells and initiate the culture immediately in order to retain the highest cell viability. 2. To thaw the cells, put the vial in 37°C water bath with gentle agitation for ~1 minute. Keep the cap out of water to minimize the risk of contamination. 3. Pipette the cells into a 15ml conical tube with 5ml fresh Endothelial Cell Medium (Cat# MD-0010). 4. Centrifuge at 1000rpm (~220g) for 5 minutes under room temperature. 5. Remove the supernatant and resuspend the cells in fresh culture medium. 6. Culture the cell in T75 flask.
Safety Precaution: it is highly recommended that protective gloves and clothing should be used when handling frozen vials.
Standard Culture Procedure 1. HPAEC can be cultured in Endothelial Cell Medium (Cat# MD-0010). 2. When cells reach ~80-90% confluence, remove the medium, and wash once with sterile PBS (5ml/T75 flask). 3. Add ~2.5 ml of 0.25% Trypsin-EDTA to the flask and incubate for ~3 minutes at 37°C. Neutralize the enzyme by adding 2-3 volumes of cell culture medium. 4. Centrifuge 1,000 rpm (~220 g) for 5 min and resuspend the cells in desired volume of medium. 1000 rpm (~220 g) 5. Seed the cells in the new culture vessels at 5 × 103 cells/cm2. |
References
[1] Meuchel LW, Thompson MA, Cassivi SD, Pabelick CM and Prakash YS. Neurotrophins induce nitric oxide generation in human pulmonary artery endothelial cells. Cardiovasc Res. 2011; 91(4): 668-676.
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