Porcine Aortic Endothelial Cell (pAEC) Isolation: An Enzyme-based Method to Isolate Endothelial Cells from Harvested Porcine Abdominal Aorta

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

1. Under aseptic conditions, remove the pig aorta from the washing buffer, and place it on a 150 mm Petri dish (Figure 1A).
2. Gently cut off the two ends of the aorta and excise excess tissue around aorta with sterile forceps and scissors again (Figure 1B). Wash the outside and inside of the aorta (over the culture dish at room temperature) with 20−50 mL of washing buffer: 1x PBS solution (pH 7....

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 por.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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#