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Isolation of endothelial progenitor cells

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https://www.readbyqxmd.com/read/28317180/anti-acsa-2-defines-a-novel-monoclonal-antibody-for-prospective-isolation-of-living-neonatal-and-adult-astrocytes
#1
Christina G Kantzer, Camille Boutin, Ina D Herzig, Carolina Wittwer, Sandy Reiß, Marie Catherine Tiveron, Jan Drewes, Thomas D Rockel, Stefanie Ohlig, Jovica Ninkovic, Harold Cremer, Sandra Pennartz, Melanie Jungblut, Andreas Bosio
Astrocytes are the most abundant cell type of the central nervous system and cover a broad range of functionalities. We report here the generation of a novel monoclonal antibody, anti-astrocyte cell surface antigen-2 (Anti-ACSA-2). Flow cytometry, immunohistochemistry and immunocytochemistry revealed that Anti-ACSA-2 reacted specifically with a not yet identified glycosylated surface molecule of murine astrocytes at all developmental stages. It did not show any labeling of non-astroglial cells such as neurons, oligodendrocytes, NG2(+) cells, microglia, endothelial cells, leukocytes, or erythrocytes...
March 20, 2017: Glia
https://www.readbyqxmd.com/read/28303468/human-very-small-embryonic-like-cells-support-vascular-maturation-and-therapeutic-revascularization-induced-by-endothelial-progenitor-cells
#2
Coralie L Guerin, Elisa Rossi, Bruno Saubamea, Audrey Cras, Virginie Mignon, Jean-Sébastien Silvestre, David M Smadja
Very small embryonic-like stem cells (VSELs) are major pluripotent stem cells defined as cells of small size being Lineage- negative, CD133-positive, and CD45-negative. We previously described that human bone marrow VSELs were able to differentiate into endothelial cells and promoted post-ischemic revascularization in mice with surgically induced critical limb ischemia. In the present work, we isolated bone marrow VSELs from patients with critical limb ischemia and studied their ability to support endothelial progenitor cells therapeutic capacity and revascularization potential...
March 16, 2017: Stem Cell Reviews
https://www.readbyqxmd.com/read/28302184/cytokine-free-directed-differentiation-of-human-pluripotent-stem-cells-efficiently-produces-hemogenic-endothelium-with-lymphoid-potential
#3
Yekaterina Galat, Svetlana Dambaeva, Irina Elcheva, Aaruni Khanolkar, Kenneth Beaman, Philip M Iannaccone, Vasiliy Galat
BACKGROUND: The robust generation of human hematopoietic progenitor cells from induced or embryonic pluripotent stem cells would be beneficial for multiple areas of research, including mechanistic studies of hematopoiesis, the development of cellular therapies for autoimmune diseases, induced transplant tolerance, anticancer immunotherapies, disease modeling, and drug/toxicity screening. Over the past years, significant progress has been made in identifying effective protocols for hematopoietic differentiation from pluripotent stem cells and understanding stages of mesodermal, endothelial, and hematopoietic specification...
March 17, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28287533/isolation-of-endothelial-progenitor-cells-from-healthy-volunteers-and-their-migratory-potential-influenced-by-serum-samples-after-cardiac-surgery
#4
Christoph Emontzpohl, David Simons, Sandra Kraemer, Andreas Goetzenich, Gernot Marx, Jürgen Bernhagen, Christian Stoppe
Endothelial progenitor cells (EPCs) are recruited from the bone marrow under pathological conditions like hypoxia and are crucially involved in the neovascularization of ischemic tissues. The origin, classification and characterization of EPCs are complex; notwithstanding, two prominent sub-types of EPCs have been established: so-called "early" EPCs (subsequently referred to as early-EPCs) and late-outgrowth EPCs (late-EPCs). They can be classified by biological properties as well as by their appearance during in vitro culture...
February 14, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28247345/isolation-culture-and-immunostaining-of-skeletal-muscle-myofibers-from-wildtype-and-nestin-gfp-mice-as-a-means-to-analyze-satellite-cell
#5
Pascal Stuelsatz, Paul Keire, Zipora Yablonka-Reuveni
Multinucleated myofibers, the functional contractile units of adult skeletal muscle, harbor mononuclear Pax7(+) myogenic progenitors on their surface between the myofiber basal lamina and plasmalemma. These progenitors, known as satellite cells, are the primary myogenic stem cells in adult muscle. This chapter describes our laboratory protocols for isolating, culturing, and immunostaining intact myofibers from mouse skeletal muscle as a means for studying satellite cell dynamics. The first protocol discusses myofiber isolation from the flexor digitorum brevis (FDB) muscle...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28244038/flow-cytometric-isolation-and-differentiation-of-adipogenic-progenitor-cells-into-brown-and-brite-beige-adipocytes
#6
Jochen Steinbring, Antonia Graja, Anne-Marie Jank, Tim J Schulz
Aside from mature adipocytes, adipose tissue harbors several distinct cell populations including immune cells, endothelial cells, and adipogenic progenitor cells (AdPCs). AdPCs represent the reservoir of regenerative cells that replenishes adipocytes during normal cellular turnover and during times of increased demand for triglyceride-storage capacity. The worldwide increase in pathologies associated with the metabolic syndrome, such as obesity and type-2 diabetes, has heightened public and scientific interest in adipose tissues and the cell biological processes of adipose tissue formation and function...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28229409/endothelial-progenitor-cells-procedure-for-cell-isolation-and-applications
#7
Garikipati V N Srikanth, Raj Kishore
Bone marrow endothelial progenitor cells (EPCs) have shown a great promise to promote ischemic tissue neovascularization and to attenuate ischemic injury in a variety of animal models, which led to EPC-based clinical trials that yielded modest but promising results. Some of the variables in the use of EPCs relate to their differential isolation and characterization protocols since the EPC literature does not identify a unique marker for these vascular progenitors. In this chapter, we present step-by-step protocols for the isolation of EPCs, their characterization and culture conditions, and their potential use in basic and clinical research...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28228349/identification-of-prdm-genes-in-human-corneal-endothelium
#8
Kostadin Rolev, G O'Donovan Dominic, Christiana Georgiou, Rajan Madhavan, Alexandra Chittka
Corneal endothelial cells (CECs) are essential for maintaining corneal stromal hydration and ensuring its transparency, which is necessary for normal vision. Dysfunction of CECs leads to stromal decompensation, loss of transparency and corneal blindness. Corneal endothelium has low proliferative potential compared to surface epithelial cells leading to poor regeneration of CEC following injury or age related decline in cellular density. The mechanisms which control proliferation and differentiation of neural crest derived CEC progenitors are yet to be clearly elucidated...
February 19, 2017: Experimental Eye Research
https://www.readbyqxmd.com/read/28223221/the-atheroma-plaque-secretome-stimulates-the-mobilization-of-endothelial-progenitor-cells-ex-vivo
#9
Francisco M Vega, Violette Gautier, Cecilia M Fernandez-Ponce, M J Extremera, A F M Altelaar, Jaime Millan, Juan C Tellez, Jose A Hernandez-Campos, Rosario Conejero, Jorge Bolivar, Ricardo Pardal, Francisco J Garcia-Cózar, Enrique Aguado, Albert J R Heck, Mª Carmen Duran-Ruiz
Endothelial progenitor cells (EPCs) constitute a promising alternative in cardiovascular regenerative medicine due to their assigned role in angiogenesis and vascular repair. In response to injury, EPCs promote vascular remodeling by replacement of damaged endothelial cells and/or by secreting angiogenic factors over the damaged tissue. Nevertheless, such mechanisms need to be further characterized. In the current approach we have evaluated the initial response of early EPCs (eEPCs) from healthy individuals after direct contact with the factors released by carotid arteries complicated with atherosclerotic plaques (AP), in order to understand the mechanisms underlying the neovascularization and remodeling properties assigned to these cells...
February 20, 2017: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/28186681/skin-derived-precursors-as-a-source-of-progenitors-for-corneal-endothelial-regeneration
#10
Emi Inagaki, Shin Hatou, Kazunari Higa, Satoru Yoshida, Shinsuke Shibata, Hideyuki Okano, Kazuo Tsubota, Shigeto Shimmura
Corneal blindness is the fourth leading cause of blindness in the world. Current treatment is allogenic corneal transplantation, which is limited by shortage of donors and immunological rejection. Skin-derived precursors (SKPs) are postnatal stem cells, which are self-renewing, multipotent precursors that can be isolated and expanded from the dermis. Facial skin may therefore be an accessible autologous source of neural crest derived cells. SKPs were isolated from facial skin of Wnt1-Cre/Floxed EGFP mouse. After inducing differentiation with medium containing retinoic acid and GSK 3-β inhibitor, SKPs formed polygonal corneal endothelial-like cells (sTECE)...
March 2017: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/28181481/dpath-software-reveals-hierarchical-haemato-endothelial-lineages-of-etv2-progenitors-based-on-single-cell-transcriptome-analysis
#11
Wuming Gong, Tara L Rasmussen, Bhairab N Singh, Naoko Koyano-Nakagawa, Wei Pan, Daniel J Garry
Developmental, stem cell and cancer biologists are interested in the molecular definition of cellular differentiation. Although single-cell RNA sequencing represents a transformational advance for global gene analyses, novel obstacles have emerged, including the computational management of dropout events, the reconstruction of biological pathways and the isolation of target cell populations. We develop an algorithm named dpath that applies the concept of metagene entropy and allows the ranking of cells based on their differentiation potential...
February 9, 2017: Nature Communications
https://www.readbyqxmd.com/read/28177697/dysfunction-of-endothelial-progenitor-cells-in-hyperlipidemic-rats-involves-the-increase-of-nadph-oxidase-derived-reactive-oxygen-species-production
#12
Ting-Bo Li, Jie-Jie Zhang, Bin Liu, Xiu-Ju Luo, Qi-Lin Ma, Jun Peng
NADPH oxidase (NOX) is a major source of reactive oxygen species (ROS) in the body and it plays a key role in mediation of oxidative injury in cardiovascular system. The purposes of this study are to evaluate the status of NOX in EPCs of hyperlipidemic rats and to determine whether NOX- derived ROS promotes the dysfunction of endothelial progenitor cells (EPCs). The rats were fed on high-fat diet for 8 weeks to establish a hyperlipidemic rat model, which showed the increased plasma lipids and the impaired functions of circulating EPCs (including the reduced abilities in migration and adhesion), accompanied by an increase in NOX activity and ROS production...
October 20, 2016: Canadian Journal of Physiology and Pharmacology
https://www.readbyqxmd.com/read/28173870/an-effective-ex-vivo-approach-for-inducing-endothelial-progenitor-cells-from-umbilical-cord-blood-cd34-cells
#13
Meng Qin, Xin Guan, Huihui Wang, Yu Zhang, Bin Shen, Qingyu Zhang, Wei Dai, Yupo Ma, Yongping Jiang
BACKGROUND: Transplantation of endothelial progenitor cells (EPCs)/endothelial cells (ECs) has been used for the treatment of ischemic diseases and hemophilia A, due to their great capacity for producing factor VIII and for repairing vascular damage. We established an effective approach to stimulate the expansion and differentiation of EPCs for potential therapeutic applications. METHODS: CD34(+) cells isolated from human cord blood were cultured in a two-step system for 21 days...
February 7, 2017: Stem Cell Research & Therapy
https://www.readbyqxmd.com/read/28173797/tumour-associated-glial-host-cells-display-a-stem-like-phenotype-with-a-distinct-gene-expression-profile-and-promote-growth-of-gbm-xenografts
#14
Lina Leiss, Ercan Mutlu, Anne Øyan, Tao Yan, Oleg Tsinkalovsky, Linda Sleire, Kjell Petersen, Mohummad Aminur Rahman, Mireille Johannessen, Sidhartha S Mitra, Hege K Jacobsen, Krishna M Talasila, Hrvoje Miletic, Inge Jonassen, Xingang Li, Nicolaas H Brons, Karl-Henning Kalland, Jian Wang, Per Øyvind Enger
BACKGROUND: Little is known about the role of glial host cells in brain tumours. However, supporting stromal cells have been shown to foster tumour growth in other cancers. METHODS: We isolated stromal cells from patient-derived glioblastoma (GBM) xenografts established in GFP-NOD/scid mice. With simultaneous removal of CD11b(+) immune and CD31(+) endothelial cells by fluorescence activated cell sorting (FACS), we obtained a population of tumour-associated glial cells, TAGs, expressing markers of terminally differentiaed glial cell types or glial progenitors...
February 7, 2017: BMC Cancer
https://www.readbyqxmd.com/read/28163178/over-expression-of-jagged1-ameliorates-aged-rat-derived-endothelial-progenitor-cell-functions-and-improves-its-transfusion-efficiency-for-rat-balloon-induced-arterial-injury
#15
Guangxu Zhu, Jinxiang Wang, Mingbao Song, Fang Zhou, Dagan Fu, Guangping Ruan, Yingying Bai, Zhengping Yu, Leilei Zhang, Xiangqing Zhu, Lan Huang, Rongqing Pang, Xinghua Pan
BACKGROUND: Endothelial progenitor cell (EPC) has significant age-dependent alterations in properties, but the role of Jagged1 in ageing-induced decline of EPC functions remains unclear. METHODS: 2 and 20 months old healthy male Sprague-Dawley rats were used in present study. Jagged1 gene transfection was performed in EPC isolated from aged (AEPC) and young rats (YEPC) respectively. Experiments were divided into four groups: a) pIRES2-EGFP (PE) group, b) PE combined N-[N-(3, 5-difluoro-phenacetyl)-1- alany1]-S-phenyglycine t-butyl ester (DAPT) (PE + D) group, c) pIRES2 EGFP- Jagged1 (PEJ) group and d) PEJ combined DAPT (PEJ+D) group...
February 2, 2017: Annals of Vascular Surgery
https://www.readbyqxmd.com/read/28150318/determine-exogenous-human-ddah2-gene-function-in-rabbit-bone-marrow-derived-endothelial-progenitor-cells-in-vitro
#16
Sara Shoeibi, Shabnam Mohammadi, Hamid Reza Sadeghnia, Elahe Mahdipour, Majid Ghayour-Mobarhan
The in vitro amplification of endothelial progenitor cells (EPCs) is an important method because of its role in gene transferring and regenerative medicine. In this study, we isolated rabbit bone marrow-derived EPCs to further manipulation and overexpression of dimethylarginine dimethylaminohydrolase (DDAH) in EPCs. Isolated EPCs were cultured, expanded in endothelial basal medium. Morphology of EPCs and expression levels of surface markers detected using immunocytochemistry staining and through the use of flow cytometery...
March 2017: Cell Biochemistry and Function
https://www.readbyqxmd.com/read/28120741/impact-of-age-on-human-adipose-stem-cells-for-bone-tissue-engineering
#17
Denis Dufrane
Bone non-union is a pathological condition in which all bone healing processes have stopped, resulting in abnormal mobility between two bone segments. The incidence of bone-related injuries will increase in an aging population, leading to such injuries reaching epidemic proportions. Tissue engineering and cell therapy using mesenchymal stem cells (MSCs) have raised the possibility of implanting living tissue for bone reconstruction. Bone marrow was first proposed as the source of stem cells for bone regeneration...
January 24, 2017: Cell Transplantation
https://www.readbyqxmd.com/read/28090691/peg3-pw1-is-a-marker-of-a-subset-of-vessel-associated-endothelial-progenitors
#18
Matteo Malinverno, Monica Corada, Luca Ferrarini, Luigi Formicola, Giovanna Marazzi, David Sassoon, Elisabetta Dejana
Vascular associated endothelial cell progenitors are still poorly studied and their role in the newly forming vasculature at embryonic or postnatal stage remains elusive. In the present work we first defined a set of genes highly expressed during embryo development and strongly down regulated in the adult mouse. In this group we then concentrated on the progenitor cell marker Peg3/PW1. By in vivo staining of the vasculature we found that only a subset of cells co-expressed endothelial markers and PW1. These cells were quite abundant in the embryo vasculature but declined in number at postnatal and adult stages...
January 16, 2017: Stem Cells
https://www.readbyqxmd.com/read/28078807/traumatized-muscle-derived-multipotent-progenitor-cells-recruit-endothelial-cells-through-vascular-endothelial-growth-factor-a-action
#19
Heidi R H Supanc, Shannon Gorman, Rocky S Tuan
Traumatized muscle, such as that debrided from blast injury sites, is considered a promising and convenient tissue source for multipotent progenitor cells (MPCs), a population of adult mesenchymal stem cell (MSC)-like cells. The present study aimed to assess the regenerative therapeutic potential of human traumatized muscle-derived MPCs, e.g., for injury repair in the blast-traumatized extremity, by comparing their pro-angiogenic potential in vitro and capillary recruitment activity in vivo to those of MSCs isolated from human bone marrow, a widely-used tissue source...
January 12, 2017: Journal of Tissue Engineering and Regenerative Medicine
https://www.readbyqxmd.com/read/28062497/increased-proangiogenic-activity-of-mobilized-cd34-progenitor-cells-of-patients-with-acute-st-segment-elevation-myocardial-infarction-role-of-differential-microrna-378-expression
#20
Christian Templin, Julia Volkmann, Maximilian Y Emmert, Pavani Mocharla, Maja Müller, Nicolle Kraenkel, Jelena-R Ghadri, Martin Meyer, Beata Styp-Rekowska, Sylvie Briand, Roland Klingenberg, Milosz Jaguszewski, Christian M Matter, Valentin Djonov, Francois Mach, Stephan Windecker, Simon P Hoerstrup, Thomas Thum, Thomas F Lüscher, Ulf Landmesser
OBJECTIVE: Proangiogenic effects of mobilized bone marrow-derived stem/progenitor cells are essential for cardiac repair after myocardial infarction. MicroRNAs (miRNA/miR) are key regulators of angiogenesis. We investigated the differential regulation of angio-miRs, that is, miRNAs regulating neovascularization, in mobilized CD34(+) progenitor cells obtained from patients with an acute ST-segment-elevation myocardial infarction (STEMI) as compared with those with stable coronary artery disease or healthy subjects...
February 2017: Arteriosclerosis, Thrombosis, and Vascular Biology
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