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Updated: Jun 21, 2026
Isolation, Expansion, and Adipogenic Induction of CD34+CD31+ Endothelial Cells from Human Omental and Subcutaneous Adipose Tissue
Published on: July 17, 2018
[The isolation, subculture and identification of human adipose derived endothelial progenitor cells]
1Department of Plastic and Reconstructive Surgery, the 9th People Hospital of Shanghai, Shanghai 200011, China.
Objective:
To investigate the methods of isolating and identifying human adipose derived EPCs.
Methods:
The cells obtained from human lipoaspirates were plated on culture dishes coated with human fibronectin and were cultured in DMEM containing 2% FBS. Cells of passage 2 cultured in EGM-2 (2% FBS) served as the induced cells (experimental group), with cells cultured in DMEM (2% FBS) as the non-induced cells (control group) . Immunofluorescence was used to detect the expression of cell markers, including CD34, vWF and PECAM-1. FACS (fluorescence activated cell sorter) was used to quantitatively analyze the expression rate of cell markers (CD34, CD45, CD133 and PECAM-1). Fluorescence microscope was used to observe the function of taking up DiI-ac-LDL by the induced cells. To determine the ability of forming capillary-like structure in three-dimensional matrices, the induced cells were also cultured in methylcellulose.
Results:
The induced cells of passage 2 exhibited cobblestone morphology, similar to that of the endothelial cells. In contrast, these morphological changes were not observed in non-induced cells. Immunofluorescence detected expression of vWF, PECAM-1 in induced cells and CD34 in non-induced cells. FACS analysis showed (67.41 +/- 13.35)% of the induced cells expressed PECAM-1 and (6.73 +/- 2.21)% of the non-induced cells expressed PECAM-1 (P < 0.01), while (72.39 +/- 13.45)% of the non-induced cells expressed CD34 and (16.06 +/- 3.86)% of the induced cells expressed CD34 (P < 0.01). Fluorescence microscopy observed the induced cells took up low-density lipoprotein (LDL). The formation of "branch-like" structure confirmed their functional activity.
Conclusion:
EPCs derived from human adipose may serve as another source of seeding cells for vascular tissue engineering.
Insights
Human adipose-derived endothelial progenitor cells (EPCs) were successfully isolated and identified. These cells show potential as a novel cell source for vascular tissue engineering applications.
Area of Science:
- Cell Biology
- Regenerative Medicine
- Biotechnology
Context:
- Endothelial progenitor cells (EPCs) are crucial for neovascularization.
- Identifying reliable sources of EPCs is vital for therapeutic applications.
- Human adipose tissue is an accessible source of stem cells.
Purpose:
- To establish and validate methods for isolating and identifying EPCs from human adipose tissue.
- To characterize the phenotype and function of adipose-derived EPCs.
- To assess their suitability for vascular tissue engineering.
Summary:
- Adipose-derived cells cultured under specific conditions (EGM-2) exhibited endothelial cell morphology and expressed key markers like vWF and PECAM-1.
- Fluorescence-activated cell sorting (FACS) confirmed significant differences in marker expression (PECAM-1, CD34) between induced and non-induced cells.
- Functional assays demonstrated the ability of these cells to uptake LDL and form capillary-like structures, confirming their endothelial progenitor characteristics.
Impact:
- Adipose-derived EPCs represent a promising and accessible cell source for vascular regeneration.
- This study provides a foundation for utilizing these cells in tissue engineering scaffolds.
- Potential applications include promoting wound healing and treating ischemic diseases.
