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Georg Siegel, Erika Fleck, Stefanie Elser, Ursula Hermanutz-Klein, Marc Waidmann, Hinnak Northoff, Erhard Seifried, Richard Schäfer
BACKGROUND: Endothelial colony-forming progenitor cells (ECFCs) are promising candidates for cell therapies. However, ECFC translation to the clinic requires optimized isolation and manufacture technologies according to good manufacturing practice (GMP). STUDY DESIGN AND METHODS: ECFCs were manufactured from steady-state peripheral blood (PB) leukapheresis (11 donors), using GMP-compliant technologies including pooled human platelet (PLT) lysate, and compared to human umbilical cord endothelial cells, human aortic endothelial cells, and human cerebral microvascular endothelial cells...
February 22, 2018: Transfusion
Kaela M Varberg, Seth Winfree, Kenneth W Dunn, Laura S Haneline
Vasculogenesis is a complex process by which endothelial stem and progenitor cells undergo de novo vessel formation. Quantitative assessment of vasculogenesis has become a central readout of endothelial progenitor cell functionality, and therefore, several attempts have been made to improve both in vitro and in vivo vasculogenesis models. However, standard methods are limited in scope, with static measurements failing to capture many aspects of this highly dynamic process. Therefore, the goal of developing this novel protocol was to assess the kinetics of in vitro vasculogenesis in order to quantitate rates of network formation and stabilization, as well as provide insight into potential mechanisms underlying vascular dysfunction...
January 31, 2018: Journal of Visualized Experiments: JoVE
Loubna Abdel Hadi, Francesca Calcaterra, Lucia Brambilla, Claudia Carenza, Giovanni Marfia, Silvia Della Bella, Laura Riboni
Endothelial colony-forming cells (ECFCs), a unique endothelial stem cell population, are highly increased in the blood of Kaposi sarcoma (KS) patients. KS-derived ECFCs (KS-ECFCs) are also endowed with increased proliferative and vasculogenic potential, thus suggesting that they may be precursors of KS spindle cells. However, the mechanisms underlying the increased proliferative activity of KS-ECFCs remain poorly understood. Sphingosine-1-phosphate (S1P) and ceramide-1-phosphate (C1P) are metabolically interconnected sphingoid mediators crucial to cell proliferation...
December 15, 2017: Journal of Leukocyte Biology
Francesco Moccia
Intracellular Ca2+ signaling drives angiogenesis and vasculogenesis by stimulating proliferation, migration, and tube formation in both vascular endothelial cells and endothelial colony forming cells (ECFCs), which represent the only endothelial precursor truly belonging to the endothelial phenotype. In addition, local Ca2+ signals at the endoplasmic reticulum (ER)-mitochondria interface regulate endothelial cell fate by stimulating survival or apoptosis depending on the extent of the mitochondrial Ca2+ increase...
January 11, 2018: International Journal of Molecular Sciences
Francesco Lodola, Umberto Laforenza, Fabio Cattaneo, Federico Alessandro Ruffinatti, Valentina Poletto, Margherita Massa, Richard Tancredi, Estella Zuccolo, Dlzar Alì Khdar, Alberto Riccardi, Marco Biggiogera, Vittorio Rosti, Germano Guerra, Francesco Moccia
Endothelial colony forming cells (ECFCs) represent a population of truly endothelial precursors that promote the angiogenic switch in solid tumors, such as breast cancer (BC). The intracellular Ca2+ toolkit, which drives the pro-angiogenic response to VEGF, is remodelled in tumor-associated ECFCs such that they are seemingly insensitive to this growth factor. This feature could underlie the relative failure of anti-VEGF therapies in cancer patients. Herein, we investigated whether and how VEGF uses Ca2+ signalling to control angiogenesis in BC-derived ECFCs (BC-ECFCs)...
November 10, 2017: Oncotarget
Jung Won Choi, Sung Min Son, Inhee Mook-Jung, Youn Joo Moon, Ji Yeoun Lee, Kyu-Chang Wang, Hyun-Seung Kang, Ji Hoon Phi, Seung Ah Choi, Sangjoon Chong, Jayoung Byun, Seung-Ki Kim
OBJECTIVE Moyamoya disease (MMD) is a unique cerebrovascular disorder characterized by the progressive occlusion of the bilateral internal carotid arteries. Endothelial colony-forming cells (ECFCs), previously termed "endothelial progenitor cells," play an important role in the pathogenesis of MMD. In this study, the authors performed morphological and functional studies of the mitochondria of ECFCs from patients with MMD to present new insights into the pathogenesis of the disease. METHODS The morphology of ECFCs from 5 MMD patients and 5 healthy controls was examined under both a transmission electron microscope and a confocal laser scanning microscope...
December 8, 2017: Journal of Neurosurgery
Emma Reid, Jasenka Guduric-Fuchs, Christina L O'Neill, Lynsey-Dawn Allen, Sarah E J Chambers, Alan W Stitt, Reinhold J Medina
Cell therapy using endothelial progenitors holds promise for vascular repair in ischemic retinopathies. Using a well-defined subpopulation of human cord blood-derived endothelial progenitors known as endothelial colony-forming cells (ECFCs), we have evaluated essential requirements for further development of this cell therapy targeting the ischemic retina, including dose response, delivery route, and toxicity. First, to evaluate therapeutic efficacy relating to cell dose, ECFCs were injected into the vitreous of mice with oxygen-induced retinopathy...
November 22, 2017: Stem Cells Translational Medicine
Estella Zuccolo, Christian Di Buduo, Francesco Lodola, Stefania Orecchioni, Giorgia Scarpellino, Dlzar Ali Kheder, Valentina Poletto, Germano Guerra, Francesco Bertolini, Alessandra Balduini, Vittorio Rosti, Francesco Moccia
Stromal cell-derived factor-1α (SDF-1α) drives endothelial colony-forming cell (ECFC) homing and incorporation within neovessels, thereby restoring tissue perfusion in ischemic tissues and favoring tumor vascularization and metastasis. SDF-1α stimulates ECFC migration by activating the Gi -protein-coupled receptor, CXCR4, and then engaging the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. Sporadic evidence showed that SDF-1α may also act through an increase in intracellular Ca2+ concentration ([Ca2+ ]i ) in bone marrow-derived hematopoietic progenitor cells and fully differentiated endothelial cells...
January 1, 2018: Stem Cells and Development
Nour C Bacha, Adeline Blandinieres, Elisa Rossi, Nicolas Gendron, Nathalie Nevo, Séverine Lecourt, Coralie L Guerin, Jean Marie Renard, Pascale Gaussem, Eduardo Angles-Cano, Chantal M Boulanger, Dominique Israel-Biet, David M Smadja
Idiopathic pulmonary fibrosis (IPF) is a devastating disease characterized by obliteration of alveolar architecture, resulting in declining lung function and ultimately death. Pathogenic mechanisms remain unclear but involve a concomitant accumulation of scar tissue together with myofibroblasts activation. Microparticles (MPs) have been investigated in several human lung diseases as possible pathogenic elements, prognosis markers and therapeutic targets. We postulated that levels and cellular origins of circulating MPs might serve as biomarkers in IPF patients and/or as active players of fibrogenesis...
November 3, 2017: Stem Cell Reviews
Ba Reun Kim, Yang Woo Kwon, Gyu Tae Park, Eun Jung Choi, Jeong Kon Seo, Il Ho Jang, Seung-Chul Kim, Hyun-Chang Ko, Sang Chul Lee, Jae Ho Kim
Angiogenic peptides have therapeutic potential for the treatment of chronic ischemic diseases. Periostin, an extracellular matrix protein expressed in injured tissues, promotes angiogenesis and tissue repair. We previously reported that in vivo administration of both recombinant full-length protein and the first FAS I domain of periostin alleviated peripheral artery occlusive disease by stimulating the migration of humane endothelial colony forming cells (ECFCs) and subsequent angiogenesis. In the present study, we ascertained the peptide sequence responsible for the periostin-induced angiogenesis...
2017: PloS One
Michelle Swearingen, Beverly Falcon, Sudhakar Chintharlapalli, Mark Uhlik
Blood vessels are crucial components for normal tissue development and homeostasis, so it is not surprising that endothelial dysfunction and dysregulation results in a variety of different pathophysiological conditions. The large number of vascular-related disorders and the emergence of angiogenesis as a major hallmark of cancer has led to significant interest in the development of drugs that target the vasculature. While several in vivo models exist to study developmental and pathological states of blood vessels, few in vitro assays have been developed that capture the significant complexity of the vascular microenvironment...
2018: Methods in Molecular Biology
Hao Zhang, Yanling Tao, Saisai Ren, Haihui Liu, Hui Zhou, Jiangwei Hu, Yongyong Tang, Bin Zhang, Hu Chen
Endothelial colony-forming cells (ECFCs) are a population of endothelial progenitor cells (EPCs) that display robust proliferative potential and vessel-forming capability. Previous studies have demonstrated that a limited number of ECFCs may be obtained from adult bone marrow, peripheral blood and umbilical cord (UC) blood. The present study describes an effective method for isolating ECFCs from human UC. The ECFCs derived from human UC displayed the full properties of EPCs. Analysis of the growth kinetics, cell cycle and colony-forming ability of the isolated human UC-ECFCs indicated that the cells demonstrated properties of stem cells, including relative stability and rapid proliferation in vitro...
November 2017: Experimental and Therapeutic Medicine
Abbas Shafiee, Jatin Patel, James S Lee, Dietmar W Hutmacher, Nicholas M Fisk, Kiarash Khosrotehrani
The clinical use of endothelial colony forming cells (ECFC) is hampered by their restricted engraftment. We aimed to assess engraftment, vasculogenic and pro-angiogenic activities of ECFC in immunocompetent (C57BL/6: WT) or immunodeficient (rag1 (-/-) C57BL/6: Rag1) mice. In addition, the impact of host immune system was investigated where ECFC were co-implanted with mesenchymal stem/stromal cells (MSC) from adult bone marrow (AdBM-MSC), fetal bone marrow (fBM-MSC), fetal placental (fPL-MSC), or maternal placental (MPL-MSC)...
October 19, 2017: Scientific Reports
Giovanna Vinci, Christophe Buffat, Stéphanie Simoncini, Farid Boubred, Isabelle Ligi, Florent Dumont, Bernard Le Bonniec, Thierry Fournier, Daniel Vaiman, Françoise Dignat-George, Umberto Simeoni
OBJECTIVE: Preterm birth is associated with altered angiogenesis and with increased risk of cardiovascular dysfunction and hypertension at adulthood. We previously demonstrated that in preterm newborns circulating cord blood endothelial progenitor cells (ECFC), responsible for angio/vasculogenesis, are reduced in number and display altered angiogenic properties. Altered angiogenic function was associated with a decreased expression of pro-angiogenic genes, among which the AMOT gene which is a strong positive regulator of angiogenesis...
2017: PloS One
Sylvain Fraineau, Carmen G Palii, Brian McNeill, Morten Ritso, William C Shelley, Nutan Prasain, Alphonse Chu, Elodie Vion, Kristy Rieck, Sharmin Nilufar, Theodore J Perkins, Michael A Rudnicki, David S Allan, Mervin C Yoder, Erik J Suuronen, Marjorie Brand
Human endothelial colony-forming cells (ECFCs) represent a promising source of adult stem cells for vascular repair, yet their regenerative capacity is limited. Here, we set out to understand the molecular mechanism restricting the repair function of ECFCs. We found that key pro-angiogenic pathways are repressed in ECFCs due to the presence of bivalent (H3K27me3/H3K4me3) epigenetic marks, which decreases the cells' regenerative potential. Importantly, ex vivo treatment with a combination of epigenetic drugs that resolves bivalent marks toward the transcriptionally active H3K4me3 state leads to the simultaneous activation of multiple pro-angiogenic signaling pathways (VEGFR, CXCR4, WNT, NOTCH, SHH)...
November 14, 2017: Stem Cell Reports
Leni Moldovan, April Barnard, Chang-Hyun Gil, Yang Lin, Maria B Grant, Mervin C Yoder, Nutan Prasain, Nicanor I Moldovan
Recently a protocol is established to obtain large quantities of human induced pluripotent stem cells (iPSC)-derived endothelial progenitors, called endothelial colony forming cells (ECFC), and of candidate smooth-muscle forming cells (SMFC). Here, the suitability for assembling in spheroids, and in larger 3D cell constructs is tested. iPSC-derived ECFC and SMFC are labeled with tdTomato and eGFP, respectively. Spheroids are formed in ultra-low adhesive wells, and their dynamic proprieties are studied by time-lapse microscopy, or by confocal microscopy...
October 14, 2017: Biotechnology Journal
Isabelle Grandvuillemin, Philippe Garrigue, Alaa Ramdani, Farid Boubred, Umberto Simeoni, Françoise Dignat-George, Florence Sabatier, Benjamin Guillet
Neonatal hypoxic-ischemic encephalopathy (NHIE) is a dramatic perinatal complication, associated with poor neurological prognosis despite neuroprotection by therapeutic hypothermia, in the absence of an available curative therapy. We evaluated and compared ready-to-use human umbilical cord blood cells (HUCBC) and bankable but allogeneic endothelial progenitors (ECFC) as cell therapy candidate for NHIE. We compared benefits of HUCBC and ECFC transplantation 48 hours after injury in male rat NHIE model, based on the Rice-Vannucci approach...
November 2017: Stem Cells Translational Medicine
Clément d'Audigier, Sophie Susen, Adeline Blandinieres, Virginie Mattot, Bruno Saubamea, Elisa Rossi, Nathalie Nevo, Séverine Lecourt, Coralie L Guerin, Blandine Dizier, Nicolas Gendron, Bertrand Caetano, Pascale Gaussem, Fabrice Soncin, David M Smadja
Egfl7 (VE-statin) is a secreted protein mostly specific to the endothelial lineage during development and in the adult and which expression is enhanced during angiogenesis. Egfl7 involvement in human postnatal vasculogenesis remains unresolved yet. Our aim was to assess Egfl7 expression in several angiogenic cell types originating from human bone marrow, peripheral blood, or cord blood. We found that only endothelial colony forming cells (ECFC), which are currently considered as the genuine endothelial precursor cells, expressed large amounts of Egfl7...
October 4, 2017: Stem Cell Reviews
Dawei Wang, Yimeng Song, Jianshu Zhang, Wei Pang, Xian Wang, Yi Zhu, Xiaoxia Li
Endothelial colony forming cells (ECFCs) were proved to take part in post-natal vasculogenesis and injury repair. The angiogenic properties of ECFCs could be influenced by various cytokines, chemokines, and growth factors. Erythropoietin (EPO) is a promising cytokine participating in angiogenesis. However, the mechanisms for EPO's proangiogenic effect still remain largely elusive. Here, we investigated the role of AMP-activated protein kinase (AMPK)-Krüppel-like factor 2 (KLF2) signaling pathway in the proangiogenic effect of EPO in ECFCs...
October 4, 2017: American Journal of Physiology. Cell Physiology
Mengyu He, Shuying Ma, Qing Cai, Yan Wu, Chengjie Shao, Hui Kong, Hong Wang, Xiaoning Zeng, Weiping Xie
Endothelial injury is considered as a trigger of pulmonary vascular lesions in the pathogenesis of hypoxic pulmonary hypertension (HPH). Although endothelial colony-forming cells (ECFCs) have vascular regeneration potential to maintain endothelial integrity, hypoxia-induced precise alteration in ECFCs function remains controversial. This study investigated the impact of hypoxia on human ECFCs function in vitro and the underlying mechanism. We found that hypoxia inhibited ECFCs proliferation, migration and angiogenesis...
September 28, 2017: Respiratory Physiology & Neurobiology
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