Porcine Aortic Endothelial Cell (pAEC) Isolation: An Enzyme-based Method to Isolate Endothelial Cells from Harvested Porcine Abdominal Aorta
Published: April 30, 2023
Abstract
Source: Zhao, Y. et al. Isolation and Culture of Primary Aortic Endothelial Cells from Miniature Pigs. J. Vis. Exp. (2019)
In this video, we demonstrate the isolation of endothelial cells from the abdominal aorta of a pig model using one-step collagenase digestion. The endothelial cells can be cultured for further downstream applications.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Isolation of Porcine Aortic Endothelial Cells Under aseptic conditions, remove the pig aorta from the washing buffer, and place it on a 150 mm Petri dish (Figure 1A). Gently cut off the two ends of the aorta and excise excess tissue around aorta with sterile forceps and scissors again (Figure 1B</stron…
Representative Results
Figure 1: Porcine aorta digestion and pAECs isolation. (A) The aorta is placed in a 150 mm Petri dish with cold washing buffer. (B) Photograph of porcine aorta after removing connective tissues. (C) The glass bar tied with a sterile surgical sutu…
Divulgaciones
The authors have nothing to disclose.
Materials
| Cell scraper | Corning | 3010# | |
| Collagenase IV | Sigma-Aldrich | C5138#-1G | |
| DMEM | Life Technologies | 11965118# | |
| Falcon 100mm Cell Culture Dish | Corning | 353003# | |
| Petri Dishes (150 x 15 mm) | Biologixgroup | 66-1515# | |
| Forceps | ShangHai medical instruments Co.,Ltd.China | ||
| Rectangular Canted Neck Cell Culture Flask with Vent Cap (T25) | Corning | 3289# | |
| Scissors | ShangHai medical instruments Co.,Ltd.China | ||
| Serological Transfer Pipettes (10ml) | JET Biofil | GSP010010# | |
| Sterile Pasteur Pipette | GeneBrick | GY0025# | |
| Surgical suture | Shanghai Pudong Jinhuan Medical Supplies Co., Ltd | 5-0# | |
| 20 x PBS solution (pH 7.4,Nuclease free) | Sangon Biotech | B540627# | |
| Penicillin/Streptomycin | Life Technologies | 15070063# |
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Porcine Aortic Endothelial Cell (pAEC) Isolation: An Enzyme-based Method to Isolate Endothelial Cells from Harvested Porcine Abdominal Aorta. J. Vis. Exp. (Pending Publication), e20926, doi: (2023).