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Published on: February 18, 2017
Comparison between total endothelial progenitor cell isolation versus enriched Cd133+ culture
Amelia Casamassimi1, Maria Luisa Balestrieri, Carmela Fiorito
1Department of General Pathology, Division of Clinical Pathology and Excellence Research Center on Cardiovascular Diseases, II University of Naples, Italy.
Journal of Biochemistry
|February 20, 2007
Summary
Isolating endothelial progenitor cells (EPCs) from enriched CD133(+) cells yields a higher percentage of functional cells compared to total peripheral blood mononuclear cells (PBMCs). Both methods show similar functional responses but differ in CD133 expression.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Immunology
Background:
- Endothelial progenitor cells (EPCs) are crucial for neovascularization in ischemic tissues.
- Accurate isolation and characterization of EPCs are vital for cellular therapies and data reproducibility.
- Current methods for EPC isolation include using total peripheral blood mononuclear cells (PBMCs) and enriched CD133(+) cells.
Purpose of the Study:
- To compare the efficacy of two EPC isolation methods: from total PBMCs versus CD133(+) enriched cells.
- To define and analyze the morphology and functional activities of EPCs obtained by each method.
- To assess the relevance of CD133 expression in intervention studies.
Main Methods:
- Comparison of EPC isolation from total PBMCs and CD133(+) enriched cells.
- Detailed analysis of cell morphology and functional activities using a novel subfield-based cell counting method.
- Assessment of cell response to TNF-alpha and glucose stimulation, including p38-phosphorylation.
- Phenotypic characterization, focusing on CD133 expression.
Main Results:
- EPCs isolated from CD133(+) enriched cells showed a significantly higher percentage (50% +/- 7%) of Dil-Ac-LDL and lectin double-positive cells compared to those from total PBMCs (23% +/- 4%).
- Both isolation methods yielded EPCs with similar responses to TNF-alpha or glucose-induced p38-phosphorylation.
- CD133(+) enriched cells exhibited significantly higher CD133 expression compared to cells from total PBMCs.
Conclusions:
- Enrichment for CD133(+) cells provides a more efficient method for isolating functional EPCs.
- While functional responses to stimuli are comparable, the increased CD133 expression in enriched cells may be relevant for therapeutic interventions.
- Standardized isolation protocols are essential for reproducible research in EPC-based therapies.
