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      Effects of obstetric factors and storage temperatures on the yield of endothelial colony forming cells from umbilical cord blood

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          Abstract

          As umbilical cord blood (UCB) is a rich source of endothelial colony-forming cells (ECFC), our aim was twofold: (1) to examine potential obstetric selection criteria for achieving the highest ECFC yields from UCB units, and (2) to determine whether transient storage temperatures of fresh UCB and cryopreservation of UCB units affected ECFC yield and function. ECFC quality was assessed before and after cryopreservation by their clonogenic proliferative potential. Of the 20 factors examined, placental weight was the only statistically significant obstetric factor that predicted ECFC frequency in UCB. Studies on the effects of storage revealed that transient storage of fresh UCB at 4°C reduced ECFC yield compared with storage at 22°C, while cryopreservation of UCB MNCs significantly reduced ECFC recoveries. To our knowledge, this is the first demonstration that placental weight and temperature of storage prior to processing or culture have significant effects on ECFC frequency in UCB. Our studies further support the evidence that cryopreservation of UCB MNCs compromises ECFC recovery.

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          Human umbilical cord blood as a potential source of transplantable hematopoietic stem/progenitor cells.

          The purpose of this study was to evaluate human umbilical cord blood as an alternative to bone marrow in the provision of transplantable stem/progenitor cells for hematopoietic reconstitution. Although no direct quantitative assay for human hematopoietic repopulating cells is at present available, the granulocyte-macrophage progenitor cell (CFU-GM) assay has been used with success as a valid indicator of engrafting capability. We examined greater than 100 collections of human umbilical cord blood for their content of nucleated cells and granulocyte-macrophage, erythroid (BFU-E), and multipotential (CFU-GEMM) progenitor cells, in many cases both before and after cryopreservation. First it was determined that granulocyte-macrophage, erythroid, and multipotential progenitor cells remained functionally viable in cord blood untreated except for addition of anticoagulant for at least 3 days at 4 degrees C or 25 degrees C (room temperature), though not at 37 degrees C, implying that these cells could be satisfactorily studied and used or cryopreserved for therapy after transport of cord blood by overnight air freight carriage from a remote obstetrical service. Granulocyte-macrophage progenitor cells from cord blood so received responded normally to stimulation by purified recombinant preparations of granulocyte-macrophage, granulocyte, and macrophage colony-stimulating factors and interleukin 3. The salient finding, based on analysis of 101 cord blood collections, is that the numbers of progenitor cells present in the low-density (less than 1.077 gm/ml) fraction after Ficoll/Hypaque separation typically fell within the range that has been reported for successful engraftment by bone marrow cells. Another observation of practical importance is that procedures to remove erythrocytes or granulocytes prior to freezing, and washing of thawed cells before plating, entailed large losses of progenitor cells, the yield of unwashed progenitor cells from unfractionated cord blood being many times greater. The provisional inference is that human umbilical cord blood from a single individual is typically a sufficient source of cells for autologous (syngeneic) and for major histocompatibility complex-matched allogeneic hematopoietic reconstitution.
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            Mesenchymal stromal cells: facilitators of successful transplantation?

            Mesenchymal stromal/stem cells (MSCs) possess immunomodulatory and reparative properties. Through specific interactions with immune cells that participate in both innate and adaptive responses, MSCs exposed to an inflammatory microenvironment can downregulate many immune effector functions. Clinical trials focusing on MSCs to treat graft-versus-host disease (GvHD) and autoimmune diseases are underway. Current analyses suggest that MSCs will improve cell and solid organ transplantation by ameliorating rejection and possibly eliminating the requirement for prolonged regimens of conventional immunosuppressive drugs. This review examines the in vitro and in vivo evidence for the clinical use of bone marrow derived MSCs. Copyright © 2010 Elsevier Inc. All rights reserved.
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              Unresolved questions, changing definitions, and novel paradigms for defining endothelial progenitor cells.

              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.
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                Author and article information

                Contributors
                +44-1865-387919 , +44-1865-763267 , kate.e.coldwell@gmail.com
                +44-1865-387919 , +44-1865-763267 , stephlee11@gmail.com
                +44-1865-387919 , +44-1865-763267 , jenniferkean@nhs.net
                +44-1865-387919 , +44-1865-763267 , Cheen.Khoo@nhsbt.nhs.uk
                +44-1865-387919 , +44-1865-763267 , grigorios.tsaknakis@gmail.com
                +44-1865-387919 , +44-1865-763267 , Jon.Smythe@nhsbt.nhs.uk
                +44-1865-387919 , +44-1865-763267 , Suzanne.Watt@nhsbt.nhs.uk
                Journal
                Angiogenesis
                Angiogenesis
                Springer Netherlands (Dordrecht )
                0969-6970
                1573-7209
                1 July 2011
                1 July 2011
                September 2011
                : 14
                : 3
                : 381-392
                Affiliations
                [1 ]Stem Cell Research Laboratory, NHS Blood and Transplant, Oxford, OX3 9BQ UK
                [2 ]Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, OX3 9DU UK
                [3 ]Medical Sciences Division, University of Melbourne, Melbourne, 3050 Australia
                [4 ]Department of Plastic and Reconstructive Surgery, Stoke Mandeville Hospital, Aylesbury, HP21 8AL UK
                Article
                9222
                10.1007/s10456-011-9222-4
                3155043
                21720855
                6c92470a-3c1d-4cb7-84d2-0005041642c6
                © The Author(s) 2011
                History
                : 3 May 2011
                : 16 June 2011
                Categories
                Original Paper
                Custom metadata
                © Springer Science+Business Media B.V. 2011

                Human biology
                cryopreservation,transient storage,endothelial colony-forming cells,umbilical cord blood,cord blood banking,endothelial progenitor cells

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