The Impact of Proliferative Potential of Umbilical Cord–Derived Endothelial Progenitor Cells and Hypoxia on Vascular Tubule Formation In Vitro

2009 ◽  
Vol 18 (2) ◽  
pp. 359-376 ◽  
Author(s):  
Youyi Zhang ◽  
Nita Fisher ◽  
Sarah E. Newey ◽  
Jon Smythe ◽  
Louise Tatton ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Umbilical Cord ◽  
Proliferative Potential ◽  
Endothelial Progenitor ◽  
Tubule Formation ◽  
The Impact

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 Low Ethanol Concentrations Promote Endothelial Progenitor Cell Capacity and Reparative Function

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Lars Brodowski ◽  
Bianca Schröder-Heurich ◽  
Berina Kipke ◽  
Cara Schmidt ◽  
Constantin S. von Kaisenberg ◽  
...  
Keyword(s):  
Cell Surface ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Cell Interaction ◽  
Cell Number ◽  
Endothelial Progenitor ◽  
Tubule Formation ◽  
Close Link

Background. Endothelial progenitor cells (EPCs) are recruited to injured endothelium and contribute to its regeneration. There is evidence that moderate ethanol consumption prevents the development and progression of atherosclerosis in a variety of in vitro and in vivo models and increases the mobilization of progenitor cells. Furthermore, there are studies that identified ethanol at low concentration as a therapeutic tool to mobilize progenitor cells in peripheral blood. At the same time, the cell number of EPCs represents a close link to cardiovascular system constitution and function and contributes to cardiovascular risk. The aim of this study was to evaluate the effect of low dose ethanol on typical features of endothelial colony-forming cells (ECFCs), a proliferative subtype of EPCs. Methods and Results. We tested whether ethanol impacts the functional abilities of ECFC (e.g., migration, tube formation, and proliferation) using in vitro assays, the intercommunication of ECFC by exploring cell surface molecules by flow cytometry, and the expression of (anti-)angiogenic molecules by ELISA. Low concentrations of ethanol concentration promoted migration, proliferation, and tubule formation of ECFC. The expression of the cell surface marker VE-cadherin, a protein which plays an important role in cell-cell interaction, was enhanced by ethanol, while (anti-)angiogenic molecule expression was not impacted. Conclusion. Ethanol at moderate concentrations increases the angiogenic abilities of endothelial progenitor cells thus possibly contributing to vasoprotection.

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

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.

Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood

Blood ◽  
2004 ◽  
Vol 104 (9) ◽  
pp. 2752-2760 ◽  
Author(s):  
David A. Ingram ◽  
Laura E. Mead ◽  
Hiromi Tanaka ◽  
Virginia Meade ◽  
Amy Fenoglio ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Disease Risk ◽  
Cardiovascular Disease Risk ◽  
Proliferative Potential ◽  
Human Umbilical Cord ◽  
Endothelial Progenitor

Abstract Emerging evidence to support the use of endothelial progenitor cells (EPCs) for angiogenic therapies or as biomarkers to assess cardiovascular disease risk and progression is compelling. However, there is no uniform definition of an EPC, which makes interpretation of these studies difficult. Although hallmarks of stem and progenitor cells are their ability to proliferate and to give rise to functional progeny, EPCs are primarily defined by the expression of cell-surface antigens. Here, using adult peripheral and umbilical cord blood, we describe an approach that identifies a novel hierarchy of EPCs based on their clonogenic and proliferative potential, analogous to the hematopoietic cell system. In fact, some EPCs form replatable colonies when deposited at the single-cell level. Using this approach, we also identify a previously unrecognized population of EPCs in cord blood that can achieve at least 100 population doublings, replate into at least secondary and tertiary colonies, and retain high levels of telomerase activity. Thus, these studies describe a clonogenic method to define a hierarchy of EPCs based on their proliferative potential, and they identify a unique population of high proliferative potential-endothelial colony-forming cells (HPP-ECFCs) in human umbilical cord blood. (Blood. 2004;104:2752-2760)

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 In Vitro and in Vivo Analysis of Endothelial Progenitor Cells from Cryopreserved Umbilical Cord Blood: Are We Ready for Clinical Application?

2010 ◽  
Vol 19 (9) ◽  
pp. 1143-1155 ◽  
Author(s):  
Valérie Vanneaux ◽  
Fida El-Ayoubi ◽  
Catherine Delmau ◽  
Catherine Driancourt ◽  
Séverine Lecourt ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Umbilical Cord ◽  
Clinical Application ◽  
Umbilical Cord Blood ◽  
Endothelial Progenitor ◽  
In Vivo Analysis

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.

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