scholarly journals Disrupted Endothelial Cell Layer and Exposed Extracellular Matrix Proteins Promote Capture of Late Outgrowth Endothelial Progenitor Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-13 ◽  
Author(s):  
Jing Zhao ◽  
Claudia-Gabriela Mitrofan ◽  
Sarah L. Appleby ◽  
Nicholas W. Morrell ◽  
Andrew M. L. Lever
Keyword(s):  
Extracellular Matrix ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Vascular Endothelial Cells ◽  
Ex Vivo ◽  
Proliferative Potential ◽  
Vascular Repair ◽  
Endothelial Progenitor ◽  
Ischaemia Reperfusion Injury

Late outgrowth endothelial progenitor cells (LO-EPC) possess a high proliferative potential, differentiate into vascular endothelial cells (EC), and form networks, suggesting they play a role in vascular repair. However, due to their scarcity in the circulation there is a requirement forex vivoexpansion before they could provide a practical cell therapy and it is currently unclear if they would home and engraft to an injury site. Using anin vitroflow system we studied LO-EPC under simulated injury conditions including EC activation, ischaemia, disrupted EC integrity, and exposed basement membrane. Perfused LO-EPC adhered to discontinuous EC paracellularly at junctional regions between adjacent cells under shear stress 0.7 dyn/cm2. The interaction was not adhesion molecule-dependent and not enhanced by EC activation. LO-EPC expressed high levels of the VE-Cadherin which may explain these findings. Ischaemia reperfusion injury decreased the interaction with LO-EPC due to cell retraction. LO-EPC interacted with exposed extracellular matrix (ECM) proteins, fibronectin and vitronectin. The interaction was mediated by integrinsα5β3,αvβ1, andαvβ3. This study has demonstrated that an injured local environment presents sufficient adhesive signals to capture flow perfused LO-EPCin vitroand that LO-EPC have properties consistent with their potential role in vascular repair.

Bovine Vessel-Derived Endothelial Cells Are Comprised of a Complete Hierarchy of Endothelial Progenitor Cells, with the Highest Proliferative Progenitors Residing in the Aorta.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3912-3912
Author(s):  
Matthew M. Harkenrider ◽  
Scott A. Johnson ◽  
Laura E. Mead ◽  
David A. Ingram ◽  
Mervin C. Yoder
Keyword(s):  
Endothelial Cells ◽  
Single Cell ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ex Vivo ◽  
Proliferative Potential ◽  
Endothelial Progenitor ◽  
Apparent Paradox ◽  
Endothelial Colony Forming Cells

Abstract Endothelial cell replication in large and small vessels is generally thought to occur at a rate of 0.1–0.6% daily. Despite this low level of cell turnover, endothelial cells derived from a variety of bovine vessels display vigorous patterns of proliferation in vitro. This apparent paradox has not been resolved to date. We have recently determined that human endothelial cells are derived through a process of endopoiesis via a hierarchy of endothelial progenitor cells (EPCs) (Blood, 2004). We have developed a single cell proliferation assay that has resolved endopoiesis into distinct stages of progenitor cell development: 1) high proliferative potential-endothelial colony forming cells (HPP-ECFC; 2001-> 10,000 cells/colony) that replate into secondary and tertiary HPP-ECFC, 2) low proliferative potential-endothelial colony forming cells (LPP-ECFC; 51–2,000 cells/colony) that form colonies greater than 50 cells but fail to replate into LPP-ECFC, 3) endothelial clusters (EC-clusters; 2–50 cells/colony) that contain fewer than 50 cells, and 4) mature differentiated endothelial cells that are non-proliferative. We hypothesized that the proliferative behavior of the bovine vessel-derived endothelial cells was due to the presence of EPCs. We purchased bovine aortic endothelial cells (BAEC), bovine pulmonary artery endothelial cells (BPAEC), and bovine coronary artery endothelial cells (BCAEC) from a commercial vendor and cultured the cells as recommended. As predicted, the endothelial cells displayed a cobblestone morphology and ingested acetylated low density lipoprotein consistent with an endothelial phenotype. We initially plated the monolayer of cells of each type at 10, 25, or 100 cells per collagen I coated 6-well tissue culture wells and determined that cells from each artery gave rise to heterogenous colony sizes with different growth potentials during a 7 day culture. We then utilized flow cytometry to single cell sort the endothelial cells of each arterial type and determined the number of cells that divided in a 14 day culture. As depicted in the TABLE, the entire hierarchy of EPCs (similar to that determined for human adult peripheral blood and umbilical cord blood) is present in the endothelial cells isolated from the bovine vessels. Of interest, our preliminary data indicate that the frequency of the most proliferative progenitors (HPP-ECFC) is higher in the BAEC than the BPAEC or BCAEC samples. These data provide a new conceptual framework for understanding the mechanisms of endothelial replacement and/or repair of aged or damaged endothelial cells. While EPCs clearly circulate, they also engraft and reside in the vessel wall. We speculate that it is the presence of these EPCs that accounts for the ability of isolated BAEC, BPAEC, and BCAEC cells to proliferate ex vivo. Single Cell Sort Colony Distributions Cell Line BAEC-1 % BAEC-2 % BCAEC % BPAEC % Mature EC 31.33 39.33 56.67 53.67 EC-clusters 2.00 2.33 10.00 5.00 LPP-ECFC 5.00 9.00 12.00 11.00 HPP-ECFC 61.67 49.33 21.33 30.33 Total colonies 68.67 60.67 43.33 46.33

Vessel wall–derived endothelial cells rapidly proliferate because they contain a complete hierarchy of endothelial progenitor cells

Blood ◽  
2005 ◽  
Vol 105 (7) ◽  
pp. 2783-2786 ◽  
Author(s):  
David A. Ingram ◽  
Laura E. Mead ◽  
Daniel B. Moore ◽  
Wayne Woodard ◽  
Amy Fenoglio ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ex Vivo ◽  
Umbilical Vein ◽  
Proliferative Potential ◽  
Endothelial Progenitor ◽  
Vessel Walls

AbstractEndothelial progenitor cells (EPCs) can be isolated from adult peripheral and umbilical cord blood and expanded exponentially ex vivo. In contrast, human umbilical vein endothelial cells (HUVECs) or human aortic endothelial cells (HAECs) derived from vessel walls are widely considered to be differentiated, mature endothelial cells (ECs). However, similar to adult- and cord blood–derived EPCs, HUVECs and HAECs derived from vessel walls can be passaged for at least 40 population doublings in vitro. Based on this paradox, we tested whether EPCs reside in HUVECs or HAECs utilizing a novel single cell deposition assay that discriminates EPCs based on their proliferative and clonogenic potential. We demonstrate that a complete hierarchy of EPCs can be identified in HUVECs and HAECs derived from vessel walls and discriminated by their clonogenic and proliferative potential. This study provides evidence that a diversity of EPCs exists in human vessels and provides a conceptual framework for determining both the origin and function of EPCs in maintaining vessel integrity.

scholarly journals Humanized large-scale expanded endothelial colony–forming cells function in vitro and in vivo

Blood ◽  
2009 ◽  
Vol 113 (26) ◽  
pp. 6716-6725 ◽  
Author(s):  
Andreas Reinisch ◽  
Nicole A. Hofmann ◽  
Anna C. Obenauf ◽  
Karl Kashofer ◽  
Eva Rohde ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Large Scale ◽  
Proliferative Potential ◽  
Vascular Networks ◽  
Endothelial Progenitor ◽  
Endothelial Colony Forming Cells ◽  
Promising Tool

Abstract Endothelial progenitor cells are critically involved in essential biologic processes, such as vascular homeostasis, regeneration, and tumor angiogenesis. Endothelial colony–forming cells (ECFCs) are endothelial progenitor cells with robust proliferative potential. Their profound vessel-forming capacity makes them a promising tool for innovative experimental, diagnostic, and therapeutic strategies. Efficient and safe methods for their isolation and expansion are presently lacking. Based on the previously established efficacy of animal serum–free large-scale clinical-grade propagation of mesenchymal stromal cells, we hypothesized that endothelial lineage cells may also be propagated efficiently following a comparable strategy. Here we demonstrate that human ECFCs can be recovered directly from unmanipulated whole blood. A novel large-scale animal protein-free humanized expansion strategy preserves the progenitor hierarchy with sustained proliferation potential of more than 30 population doublings. By applying large-scale propagated ECFCs in various test systems, we observed vascular networks in vitro and perfused vessels in vivo. After large-scale expansion and cryopreservation phenotype, function, proliferation, and genomic stability were maintained. For the first time, proliferative, functional, and storable ECFCs propagated under humanized conditions can be explored in terms of their therapeutic applicability and risk profile.

scholarly journals Role of β2-integrins for homing and neovascularization capacity of endothelial progenitor cells

2004 ◽  
Vol 201 (1) ◽  
pp. 63-72 ◽  
Author(s):  
Emmanouil Chavakis ◽  
Alexandra Aicher ◽  
Christopher Heeschen ◽  
Ken-ichiro Sasaki ◽  
Ralf Kaiser ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ex Vivo ◽  
Limb Ischemia ◽  
Endothelial Progenitor ◽  
Cell Monolayers ◽  
Β2 Integrins

The mechanisms of homing of endothelial progenitor cells (EPCs) to sites of ischemia are unclear. Here, we demonstrate that ex vivo–expanded EPCs as well as murine hematopoietic Sca-1+/Lin− progenitor cells express β2-integrins, which mediate the adhesion of EPCs to endothelial cell monolayers and their chemokine-induced transendothelial migration in vitro. In a murine model of hind limb ischemia, Sca-1+/Lin− hematopoietic progenitor cells from β2-integrin–deficient mice are less capable of homing to sites of ischemia and of improving neovascularization. Preactivation of the β2-integrins expressed on EPCs by activating antibodies augments the EPC-induced neovascularization in vivo. These results provide evidence for a novel function of β2-integrins in postnatal vasculogenesis.

Unresolved questions, changing definitions, and novel paradigms for defining endothelial progenitor cells

Blood ◽  
2005 ◽  
Vol 106 (5) ◽  
pp. 1525-1531 ◽  
Author(s):  
David A. Ingram ◽  
Noel M. Caplice ◽  
Mervin C. Yoder
Keyword(s):  
Endothelial Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Circulating Endothelial Cells ◽  
Proliferative Potential ◽  
Endothelial Progenitor ◽  
Detailed Understanding ◽  
Circulating Endothelial Progenitor Cells

Abstract The field of vascular biology has been stimulated by the concept that circulating endothelial progenitor cells (EPCs) may play a role in neoangiogenesis (postnatal vasculogenesis). One problem for the field has been the difficulty in accurately defining an EPC. Likewise, circulating endothelial cells (CECs) are not well defined. The lack of a detailed understanding of the proliferative potential of EPCs and CECs has contributed to the controversy in identifying these cells and understanding their biology in vitro or in vivo. A novel paradigm using proliferative potential as one defining aspect of EPC biology suggests that a hierarchy of EPCs exists in human blood and blood vessels. The potential implications of this view in relation to current EPC definitions are discussed.

scholarly journals Particle Radiation-Induced Nontargeted Effects in Bone-Marrow-Derived Endothelial Progenitor Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-15 ◽  
Author(s):  
Sharath P. Sasi ◽  
Daniel Park ◽  
Sujatha Muralidharan ◽  
Justin Wage ◽  
Albert Kiladjian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ex Vivo ◽  
Fold Increase ◽  
Endothelial Progenitor ◽  
Particle Radiation ◽  
Cytokines And Chemokines ◽  
Radiation Induced

Bone-marrow- (BM-) derived endothelial progenitor cells (EPCs) are critical for endothelial cell maintenance and repair. During future space exploration missions astronauts will be exposed to space irradiation (IR) composed of a spectrum of low-fluence protons (1H) and high charge and energy (HZE) nuclei (e.g., iron-56Fe) for extended time. How the space-type IR affects BM-EPCs is limited. In media transfer experimentsin vitrowe studied nontargeted effects induced by1H- and56Fe-IR conditioned medium (CM), which showed significant increase in the number of p-H2AX foci in nonirradiated EPCs between 2 and 24 h. A 2–15-fold increase in the levels of various cytokines and chemokines was observed in both types of IR-CM at 24 h.Ex vivoanalysis of BM-EPCs from single, low-dose, full-body1H- and56Fe-IR mice demonstrated a cyclical (early 5–24 h and delayed 28 days) increase in apoptosis. This early increase in BM-EPC apoptosis may be the effect of direct IR exposure, whereas late increase in apoptosis could be a result of nontargeted effects (NTE) in the cells that were not traversed by IR directly. Identifying the role of specific cytokines responsible for IR-induced NTE and inhibiting such NTE may prevent long-term and cyclical loss of stem and progenitors cells in the BM milieu.

In Vitro Angiogenesis Properties of Endothelial Progenitor Cells: A Promising Tool for Vascularization of Ex Vivo Engineered Tissues

2007 ◽  
Vol 13 (7) ◽  
pp. 1413-1420 ◽  
Author(s):  
Günter Finkenzeller ◽  
Nestor Torio-Padron ◽  
Arash Momeni ◽  
Alexander T. Mehlhorn ◽  
G. Björn Stark
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Ex Vivo ◽  
Endothelial Progenitor ◽  
Promising Tool ◽  
Engineered Tissues ◽  
In Vitro Angiogenesis

scholarly journals Cilostazol Improves Proangiogenesis Functions in Human Early Endothelial Progenitor Cells through the Stromal Cell-Derived Factor System and Hybrid Therapy Provides a Synergistic EffectIn Vivo

2016 ◽  
Vol 2016 ◽  
pp. 1-18 ◽  
Author(s):  
Shih-Ya Tseng ◽  
Ting-Hsing Chao ◽  
Yi-Heng Li ◽  
Chung-Lung Cho
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Stromal Cell ◽  
Ex Vivo ◽  
Akt Signaling ◽  
Endothelial Progenitor ◽  
Hybrid Therapy ◽  
Stromal Cell Derived Factor

This study investigated the effect of cilostazol on proangiogenesis functions in human early endothelial progenitor cells (EPCs)in vitroand the therapeutic implication of hybrid therapy with cilostazol and human early EPCsin vivo. Cilostazol significantly increased colony-forming units and enhanced differentiation of EPCs toward endothelial lineage. Treatments resulted in antiapoptotic effects and stimulated proliferation and migration andin vitrovascular tube formation through activation of stromal cell-derived factor-1 (SDF-1)/C-X-C chemokine receptor type 4 (CXCR4)/phosphatidylinositol-3 kinase (PI3K)/Akt signaling pathway. Blood flow recovery and capillary density in murine ischemic hindlimbs were significantly improved in cilostazol-treated, human early EPCs-treated, and cotreatment groups. The effects were attenuated with SDF-1αinhibition. Plasma SDF-1αlevels were significantly higher in 3 active treatment groups after surgery, with greatest effects observed in hybrid therapy. The angiogenic effects of transplanted EPCs pretreated with cilostazolex vivowere superior to untreated EPCs usingin vivoMatrigel assay. Implanted EPCs were incorporated into the capillary, with pretreatment or cotreatment with cilostazol resulting in enhanced effects. Taken together, cilostazol promotes a large number of proangiogenic functions in human early EPCs through activation of SDF-1/CXCR4/PI3K/Akt signaling, and hybrid therapy provides a synergistic effectin vivo. Cotreatment may be beneficial in ischemic disease.

scholarly journals Enhanced Adhesion of Early Endothelial Progenitor Cells to Radiation-induced Senescence-like Vascular Endothelial Cells in vitro

2009 ◽  
Vol 50 (5) ◽  
pp. 469-475 ◽  
Author(s):  
Nuttawut SERMSATHANASAWADI ◽  
Hideto ISHII ◽  
Kaori IGARASHI ◽  
Masahiko MIURA ◽  
Masayuki YOSHIDA ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial ◽  
Endothelial Progenitor ◽  
Radiation Induced
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