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Updated: May 1, 2026
Isolation and Primary Culture of Mouse Aortic Endothelial Cells
Published on: December 19, 2016
Development of immortalized mouse aortic endothelial cell lines
Chih-Wen Ni, Sandeep Kumar, Casey J Ankeny
1Wallace H, Coulter Department of Biomedical Engineering Georgia Institute of Technology and Emory University, 1760 Haygood Drive, Health Science Research Building, E-170, Atlanta, GA 30322, USA. hanjoong.jo@bme.gatech.edu.
Background:
The understanding of endothelial cell biology has been facilitated by the availability of primary endothelial cell cultures from a variety of sites and species; however, the isolation and maintenance of primary mouse aortic endothelial cells (MAECs) remain a formidable challenge. Culturing MAECs is difficult as they are prone to phenotypic drift during culture. Therefore, there is a need to have a dependable in vitro culture system, wherein the primary endothelial cells retain their properties and phenotypes.
Methods:
Here, we developed an effective method to prepare immortalized MAEC (iMAEC) lines. Primary MAECs, initially isolated from aortic explants, were immortalized using a retrovirus expressing polyoma middle T-antigen. Immortalized cells were then incubated with DiI-acetylated-low density lipoprotein and sorted via flow cytometry to isolate iMAECs.
Results:
iMAECs expressed common markers of endothelial cells, including PECAM1, eNOS, VE-cadherin, and von Willebrand Factor. iMAECs aligned in the direction of imposed laminar shear and retained the ability to form tubes. Using this method, we have generated iMAEC lines from wild-type and various genetically modified mice such as p47phox-/-, eNOS-/-, and caveolin-1-/-.
Conclusion:
In summary, generation of iMAEC lines from various genetically modified mouse lines provides an invaluable tool to study vascular biology and pathophysiology.
Insights
Researchers developed a new method to create immortalized mouse aortic endothelial cells (iMAECs) that retain their properties. This advance provides a reliable tool for studying vascular biology and disease.
Area of Science:
- Vascular Biology
- Endothelial Cell Biology
- Cell Culture
Background:
- Primary mouse aortic endothelial cells (MAECs) are crucial for research but difficult to culture due to phenotypic drift.
- Existing methods for culturing MAECs present challenges in maintaining cell properties.
- A dependable in vitro system for MAECs is needed to preserve their phenotypes.
Purpose of the Study:
- To develop an effective method for preparing immortalized MAEC (iMAEC) lines.
- To establish a stable cell culture system for MAECs.
- To facilitate research in vascular biology and pathophysiology.
Main Methods:
- Primary MAECs were isolated from aortic explants.
- Retroviral expression of polyoma middle T-antigen was used for immortalization.
- DiI-acetylated-low density lipoprotein incubation and flow cytometry sorting were employed to isolate iMAECs.
Main Results:
- Generated immortalized MAEC (iMAEC) lines express key endothelial markers (PECAM1, eNOS, VE-cadherin, von Willebrand Factor).
- iMAECs demonstrated characteristic endothelial functions, including alignment under laminar shear and tube formation.
- iMAEC lines were successfully derived from wild-type and genetically modified mouse models.
Conclusions:
- The generation of iMAEC lines offers a valuable tool for studying vascular biology.
- This method provides a stable in vitro model for investigating vascular pathophysiology.
- The availability of iMAECs from various mouse models aids in studying genetic influences on vascular function.
