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Updated: Aug 7, 2026
Isolation of Endothelial Progenitor Cells from Human Umbilical Cord Blood
Published on: September 14, 2017
Adult endothelial progenitor cells from human peripheral blood maintain monocyte/macrophage function throughout in
Shi Ju Zhang1, Hao Zhang, Ying Jie Wei
1Research Center for Cardiovascular Regenerative Medicine, Ministry of Health, West District, Beijing 100037, China.
Abstract:
Mononuclear cells (MNCs) isolated from peripheral blood by density gradient centrifugation were plated on human fibronectin-coated culture plates and cultured in EGM-2 medium. Attached spindle-shaped cells, reported as endothelial progenitor cells (EPCs) by some investigators, had elongated from adherent round cells, but had not proliferated from a small number of cells as supposed previously. The growth curve of the primary EPCs showed that the cells had little proliferative capacity. Flow cytometry analysis showed that the cells could express some of the endothelial lineage markers, while they could also express CD14, which is considered a marker of monocyte/macrophage lineages throughout culture. In endothelial function assays, the cells demonstrated a lower level of expression of eNOS than mature endothelial cells in the reverse transcription-polymerase chain reaction and did not show an ability to develop tube-like structures in angiogenesis assay in vitro. In this study, we identified the monocytoid function of EPCs by the combined Dil-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and Indian ink uptake tests. All the cells were double positive for Dil-Ac-LDL and Indian ink uptake at days 4, 14 and 28 of culture, which means the EPCs maintained monocytoid function throughout the culture. Therefore, although adult EPCs from peripheral MNCs have some endothelial lineage properties, they maintain typical monocytic function and have little proliferative capacity.
Insights
Adult endothelial progenitor cells (EPCs) from peripheral blood show limited proliferation and endothelial function. These cells primarily exhibit monocytic characteristics, challenging their role as true EPCs.
Area of Science:
- Cell Biology
- Hematology
- Immunology
Background:
- Endothelial progenitor cells (EPCs) are crucial for vascular repair.
- The origin and precise function of adult EPCs remain debated.
- Investigating EPCs' characteristics is vital for regenerative medicine.
Purpose of the Study:
- To characterize the proliferative capacity and functional attributes of adult EPCs.
- To determine if peripheral blood-derived EPCs possess true endothelial lineage properties.
- To elucidate the functional phenotype of cells identified as EPCs.
Main Methods:
- Isolation and culture of mononuclear cells (MNCs) from peripheral blood.
- Assessment of cell proliferation using growth curves.
- Flow cytometry for lineage marker expression (endothelial and monocytic markers).
- Endothelial function assays (eNOS expression, in vitro angiogenesis).
- Dil-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and Indian ink uptake assays.
Main Results:
- Cultured cells exhibited limited proliferation, contrary to previous assumptions.
- Cells expressed some endothelial markers but also CD14, a monocyte marker.
- Low eNOS expression and lack of tube formation indicated impaired endothelial function.
- Consistent positive staining for Dil-Ac-LDL and Indian ink confirmed sustained monocytic function.
Conclusions:
- Adult peripheral blood EPCs possess limited proliferative capacity.
- These cells display a mixed phenotype with significant monocytic characteristics.
- Their functional assays suggest they are not fully committed endothelial cells, but rather retain monocytic functions.
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