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Updated: May 5, 2026
Isolation of Embryonic Ventricular Endothelial Cells
Published on: July 20, 2013
Isolation, characterization, and transplantation of cardiac endothelial cells
Busadee Pratumvinit1, Kanit Reesukumal, Kajohnkiart Janebodin
1Department of Pathology, Institute for Stem Cell and Regenerative Medicine, School of Medicine, University of Washington, Seattle, WA 98109, USA ; Department of Clinical Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Insights
Researchers developed a new flow cytometry method to isolate and expand cardiac endothelial cells (CECs). This technique enables better study of CEC biology and applications in therapeutic angiogenesis.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Regenerative Medicine
Background:
- Isolating and expanding cardiac endothelial cells (CECs) for research and clinical use is challenging due to difficulties in obtaining pure cell populations and maintaining them in culture.
- Existing methods lack specific markers for myocardial endothelial cells and suitable long-term culture conditions.
Purpose of the Study:
- To develop an effective method for the isolation, characterization, and ex vivo expansion of cardiac endothelial cells from murine hearts.
- To enable further study of endothelial cell biology and explore clinical applications like neoangiogenesis.
Main Methods:
- Murine hearts were dissociated using mechanical and enzymatic methods.
- Flow cytometry with CD31 and Sca-1 markers was used to isolate endothelial cells, excluding CD45+ hematopoietic cells.
- In vitro functional assays (tube formation, acetylated-LDL uptake) and in vivo matrigel plug assays were performed.
Main Results:
- A multicolor flow cytometry method successfully isolated CD31+/Sca-1+ cardiac endothelial cells.
- Atrial-derived CECs exhibited faster growth than ventricular-derived CECs.
- Isolated CECs maintained key endothelial functions in vitro and formed microvessels in vivo, demonstrating engraftment capacity.
Conclusions:
- The developed multicolor flow cytometry method provides an effective means to isolate and purify cardiac endothelial cells from murine hearts.
- Ex vivo expanded CECs can be utilized for detailed biological studies and hold potential for clinical applications in therapeutic angiogenesis.
Abstract:
Isolation and ex vivo expansion of cardiac endothelial cells have been a recurrent challenge due to difficulties in isolation, cell heterogeneity, lack of specific markers to identify myocardial endothelial cells, and inadequate conditions to maintain long-term cultures. Herein, we developed a method for isolation, characterization, and expansion of cardiac endothelial cells applicable to study endothelial cell biology and clinical applications such as neoangiogenesis. First, we dissociated the cells from murine heart by mechanical disaggregation and enzymatic digestion. Then, we used flow cytometry coupled with specific markers to isolate endothelial cells from murine hearts. CD45+ cells were gated out to eliminate the hematopoietic cells. CD31+/Sca-1+ cells were isolated as endothelial cells. Cells isolated from atrium grew faster than those from ventricle. Cardiac endothelial cells maintain endothelial cell function such as vascular tube formation and acetylated-LDL uptake in vitro. Finally, cardiac endothelial cells formed microvessels in dorsal matrigel plug and engrafted in cardiac microvessels following intravenous and intra-arterial injections. In conclusion, our multicolor flow cytometry method is an effective method to analyze and purify endothelial cells from murine heart, which in turn can be ex vivo expanded to study the biology of endothelial cells or for clinical applications such as therapeutic angiogenesis.
