Ruixiang  Li,Fanxing Meng,Yonggang  Zhao,
A Simple and Convenient Method to Isolate and Culture Rat Aorta Endothelial Cells
Journal of Integrative Cardiology Open Access
2021
2674-2489
http://dx.doi.org/10.31487/j.JICOA.2021.01.03
https://www.sciencerepository.org/a-simple-and-convenient-method-to-isolate-and-culture_JICOA-2021-1-103 
Abstract: Aim: Endothelial injury results in vascular disorders. Culture of endothelial cells (ECs) is important to
investigate the pathogenesis of vascular diseases. ECs of rat aorta are widely applied in these researches.
However, there are many problems to isolate them, which make it hard to culture primary ECs. In the present
study, we established a simple and convenient method to isolate and culture rat aorta ECs.
Methods: Two 5ml centrifuge tubes were prepared. An opening (3 mm × 25 mm) was made in one tube
wall while the other was cut in half longitudinally. After the rat aorta was dissected along one side and
attached to the wall of the half tube, the tube with an opening and the half tube with aorta were mated and
secured by a rotund metal clip. Then, ECs were digested via the opening by 0.2% collagenase I at 37℃ for
1 h and identified by immunofluorescence of von Willebrand factor (vWF) and smooth muscle actin (SMA).
Results: A simple method for isolating ECs were established. Cultured ECs exhibited typical “cobblestone”
morphology at confluence. vWF was positively expressed in the ECs, and neither the negative stain of vWF
nor the positive stain of SMA was found.
Conclusion: This simple and convenient method to isolate and culture ECs of rat aorta can be
implemented with readily available 5ml centrifuge tubes and will greatly benefit the research of
vascular diseases.Keywords: Endothelial cell, rat, aorta, smooth muscle cell, in vitro