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Updated: May 26, 2026
Isolation of Endothelial Progenitor Cells from Human Umbilical Cord Blood
Published on: September 14, 2017
Improved culture-based isolation of differentiating endothelial progenitor cells from mouse bone marrow mononuclear
Haruki Sekiguchi1, Masaaki Ii, Kentaro Jujo
1Group of Vascular Regeneration Research, Institute of Biomedical Research and Innovation, RIKEN Center for Developmental Biology, Kobe, Japan.
Abstract:
Numerous endothelial progenitor cell (EPC)-related investigations have been performed in mouse experiments. However, defined characteristics of mouse cultured EPC have not been examined. We focused on fast versus slow adherent cell population in bone marrow mononuclear cells (BMMNCs) in culture and examined their characteristics. After 24 h-culture of BMMNCs, attached (AT) cells and floating (FL) cells were further cultured in endothelial differentiation medium separately. Immunological and molecular analyses exhibited more endothelial-like and less monocyte/macrophage-like characteristics in FL cells compared with AT cells. FL cells formed thick/stable tube and hypoxia or shear stress overload further enhanced these endothelial-like features with increased angiogenic cytokine/growth factor mRNA expressions. Finally, FL cells exhibited therapeutic potential in a mouse myocardial infarction model showing the specific local recruitment to ischemic border zone and tissue preservation. These findings suggest that slow adherent (FL) but not fast attached (AT) BMMNCs in culture are EPC-rich population in mouse.
Insights
Slow-adhering cells from mouse bone marrow mononuclear cells (BMMNCs) are rich in endothelial progenitor cells (EPCs). These EPCs demonstrate enhanced endothelial characteristics and therapeutic potential in myocardial infarction models.
Area of Science:
- Cardiovascular Biology
- Stem Cell Research
- Regenerative Medicine
Background:
- Endothelial progenitor cells (EPCs) are crucial for vascular repair.
- Previous studies on mouse EPCs lack detailed characterization of cultured populations.
- Understanding mouse EPC heterogeneity is vital for translational research.
Purpose of the Study:
- To characterize distinct cell populations derived from mouse bone marrow mononuclear cells (BMMNCs) in culture.
- To identify which adherent fraction (fast vs. slow) represents a more potent EPC population.
- To evaluate the therapeutic potential of identified EPCs in a preclinical model.
Main Methods:
- Isolation and differential culture of fast-attached (AT) and floating (FL) cells from BMMNCs.
- Immunological and molecular analyses to assess endothelial and myeloid markers.
- In vitro tube formation assays under normoxia and stress conditions (hypoxia, shear stress).
- Assessment of angiogenic gene expression.
- In vivo evaluation in a mouse myocardial infarction model.
Main Results:
- Floating (FL) cells exhibited significantly more endothelial-like characteristics and fewer monocyte/macrophage markers compared to attached (AT) cells.
- FL cells demonstrated superior in vitro tube formation capacity, which was further enhanced by hypoxia and shear stress.
- FL cells showed increased expression of angiogenic cytokine and growth factor mRNAs.
- In vivo, FL cells were recruited to the ischemic border zone in a myocardial infarction model, preserving cardiac tissue.
Conclusions:
- Slow-adherent (FL) BMMNCs, not fast-adherent (AT) cells, represent the primary EPC-rich population in mouse bone marrow cultures.
- These identified EPCs possess robust endothelial-forming capabilities and therapeutic efficacy in ischemic heart disease.
- This study provides a refined method for isolating and characterizing functional mouse EPCs for further investigation.
