Abstract
Background aims
Human endothelial progenitor cells (EPC) play an important role in regenerative medicine
and contribute to neovascularization on vessel injury. They are usually enriched from
peripheral blood, cord blood and bone marrow. In human fat tissue, EPC are rare and
their isolation remains a challenge.
Methods
Fat tissue was prepared by collagenase digestion, and the expression of specific marker
proteins was evaluated by flow cytometry in the stromal vascular fraction (SVF). For
enrichment, magnetic cell sorting was performed with the use of CD133 microbeads and
EPC were cultured until colonies appeared. A second purification was performed with
CD34; additional isolation steps were performed with the use of a combination of CD34
and CD31 microbeads. Enriched cells were investigated by flow cytometry for the expression
of endothelial specific markers, by Matrigel assay and by the uptake of acetylated
low-density lipoprotein.
Results
The expression pattern confirmed the heterogeneous nature of the SVF, with rare numbers
of CD133+ detectable. EPC gained from the SVF by magnetic enrichment showed cobblestone
morphology of outgrowth endothelial cells and expressed the specific markers CD31,
CD144, vascular endothelial growth factor (VEGF)R2, CD146, CD73 and CD105. Functional
integrity was confirmed by uptake of acetylated low-density lipoprotein and the formation
of tube-like structures on Matrigel.
Conclusions
Rare EPC can be enriched from human fat tissue by magnetic cell sorting with the use
of a combination of microbeads directed against CD133, an early EPC marker, CD34,
a stem cell marker, and CD31, a typical marker for endothelial cells. In culture,
they differentiate into EC and hence could have the potential to contribute to neovascularization
in regenerative medicine.
Key Words
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References
- Endothelial precursors in vascular repair.Microvasc Res. 2010; 79: 193-199
- Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration.Nat Med. 2003; 9: 702-712
- Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization.Proc Natl Acad Sci U S A. 2000; 97: 3422-3427
- Molecular analysis of endothelial progenitor cell (EPC) subtypes reveals two distinct cell populations with different identities.BMC Med Genomics. 2010; 3: 18
- Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis.Arterioscler Thromb Vasc Biol. 2004; 24: 288-293
- Retention of a differentiated endothelial phenotype by outgrowth endothelial cells isolated from human peripheral blood and expanded in long-term cultures.Cell Tissue Res. 2006; 326: 79-92
- In vitro differentiation of endothelial cells from AC133-positive progenitor cells.Blood. 2000; 95: 3106-3112
- Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors.Blood. 2000; 95: 952-958
- CD34 expression is regulated reciprocally with adhesion molecules in vascular endothelial cells in vitro.Blood. 1993; 81: 1001-1008
- Stromal vascular progenitors in adult human adipose tissue.Cytometry A. 2010; 77: 22-30
- Isolation of putative progenitor endothelial cells for angiogenesis.Science. 1997; 275: 964-967
- Post-natal endothelial progenitor cells for neovascularization in tissue regeneration.Cardiovasc Res. 2003; 58: 390-398
- Different expression of NOS isoforms in early endothelial progenitor cells derived from peripheral and cord blood.J Cell Biochem. 2007; 102: 992-1001
- Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood.Blood. 2004; 104: 2752-2760
- Are purified or expanded cord blood-derived CD133+ cells better at improving cardiac function?.Exp Biol Med. 2010; 235: 119-129
- Human adipose tissue-derived stem cells differentiate into endothelial cells in vitro and improve postnatal neovascularization in vivo.Biochem Biophys Res Commun. 2005; 332: 370-379
- Induction of mesenchymal to endothelial transformation of adipose-derived stem cells.J Heart Valve Dis. 2010; 19: 736-744
- Functional endothelial progenitor cells derived from adipose tissue show beneficial effect on cell therapy of traumatic brain injury.Neurosci Lett. 2010; 473: 186-191
- Plasticity of human adipose lineage cells toward endothelial cells: physiological and therapeutic perspectives.Circulation. 2004; 109: 656-663
- Adipogenic potential of adipose stem cell subpopulations.Plast Reconstr Surg. 2011; 128: 663-672
- Isolation and transcription profiling of purified uncultured human stromal stem cells: alteration of gene expression after in vitro cell culture.Mol Biol Cell. 2005; 16: 1131-1141
- Three-dimensional perfusion culture of human adipose tissue-derived endothelial and osteoblastic progenitors generates osteogenic constructs with intrinsic vascularization capacity.Stem Cells. 2007; 25: 1823-1829
- Human CD34/CD90 ASCs are capable of growing as sphere clusters, producing high levels of VEGF and forming capillaries.PloS One. 2009; 4: e6537
- Preparation and characterization of culture of CD146+ cells from human adipose tissue.Bull Exp Biol Med. 2010; 149: 113-118
- Human adipose tissue as a source of cells with angiogenic potential.Cell Transplant. 2012; 21: 235-250
- Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates.J Cell Physiol. 2006; 208: 64-76
- Improvement of postnatal neovascularization by human adipose tissue-derived stem cells.Circulation. 2004; 110: 349-355
- Human adipose tissue endothelial cells promote preadipocyte proliferation.Am J Physiol Endocrinol Metab. 2001; 281: E1037-E1044
- A familiar stranger: CD34 expression and putative functions in SVF cells of adipose tissue.World J Stem Cells. 2013; 5: 1-8
- Dose-dependent immunomodulatory effect of human stem cells from amniotic membrane: a comparison with human mesenchymal stem cells from adipose tissue.Tissue Eng. 2007; 13: 1173-1183
- Autologous bone-marrow stem-cell transplantation for myocardial regeneration.Lancet. 2003; 361: 45-46
- Outgrowth endothelial cells: characterization and their potential for reversing ischemic retinopathy.Invest Ophthalmol Vis Sci. 2010; 51: 5906-5913
- Endothelial progenitor cells as a therapeutic option in peripheral arterial disease.Eur J Vasc Endovasc Surg. 2009; 38: 475-481
- Treating fat grafts with human endothelial progenitor cells promotes their vascularization and improves their survival in diabetes mellitus.Plast Reconstr Surg. 2012; 130: 801-811
- Concerted regulation of CD34 and CD105 accompanies mesenchymal stromal cell derivation from human adventitial stromal cell.Stem Cells Dev. 2012; 22: 815-827
- Mesenchymal markers on human adipose stem/progenitor cells.Cytometry A. 2013; 83: 134-140
- In vitro neovasculogenic potential of resident adipose tissue precursors.Am J Physiol Cell Physiol. 2008; 295: C1271-C1280
- Angiogenic potential of endothelial progenitor cells and embryonic stem cells.Vasc Cell. 2011; 3: 11
- Endothelial nitric oxide synthase as a marker for human endothelial progenitor cells.Tohoku J Exp Med. 2010; 221: 19-27
- Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry.Cytometry B Clin Cytom. 2005; 64: 1-8
Holnthoner W, Hohenegger K, Husa A, Muehleder S, Meinl A, Peterbauer-Scherb A, et al. Adipose-derived stem cells induce vascular tube formation of outgrowth endothelial cells in a fibrin matrix [published online ahead of print October 5, 2012]. J Tissue Eng Regen Med. http://dx.doi.org/10.1002/term.1620.
- A comparison of the tube forming potentials of early and late endothelial progenitor cells.Exp Cell Res. 2008; 314: 430-440
- Robust functional vascular network formation in vivo by cooperation of adipose progenitor and endothelial cells.Circ Res. 2009; 104: 1410-1420
- Adipose tissue progenitor cells directly interact with endothelial cells to induce vascular network formation.Tissue Eng Part A. 2010; 16: 2953-2966
- Human adipose-derived stem cells enhance the angiogenic potential of endothelial progenitor cells, but not of human umbilical vein endothelial cells.Tissue Eng Part A. 2013; 19: 166-174
Article info
Publication history
Accepted:
June 29,
2013
Received:
March 22,
2013
Identification
Copyright
© 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
