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High Yield Recovery of Equine Mesenchymal Stem Cells from Umbilical Cord Matrix/Wharton’s Jelly Using a Semi-automated Process

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Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1235))

Abstract

Umbilical cord is an abundant source of perinatal, plastic adherent mesenchymal stem cells (UC-MSCs). UC-MSCs exhibit robust stemness and strong immunosuppressive and regenerative effects in vivo. This protocol describes enzymatic and mechanical dissociation of umbilical cord matrix (Wharton’s jelly) that results in efficient isolation of large numbers of fresh nucleated umbilical cord regenerative cells (UC-RCs) that, when cultured on plastic, exhibit similar characteristics of UC-MSCs. This protocol potentially alleviates the need for culture expansion to obtain large numbers of cells required for clinical application. Dissociation is achieved with a blend of collagenase and neutral proteases with agitation at 37 °C in a semi-automatic system. Average expected yield is 1.65 × 106 cells/g tissue with 93 % viability. This protocol has been successfully used to isolate an uncultured nucleated regenerative cell population (also referred to as stromal vascular fraction or SVF) from surgically debrided skin and from human, equine, and canine adipose tissue. The procedure requires less than 30 min for tissue dissection and less than 100 min for cell extraction. Quickly obtaining a large number of UC-RCs that have pluripotent differentiation capacity without the complexity and risks of culture expansion could simplify and expand the use of UC-RCs in clinical as well as research applications.

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Author notes

    Authors and Affiliations

    1. Department of Surgery, Tulane University, 1430 Tulane Abenue, New Orleans, LA, USA

      Timo Z. Nazari-Shafti, Eckhard U. Alt & Scott R. McClure

    2. InGeneron, Inc., Houston, TX, USA

      Ivone G. Bruno, Rudy F. Martinez & Michael E. Coleman

    3. Department of Veterinary Clinical Sciences, Iowa State University, Ames, IA, USA

      Scott R. McClure

    Authors
    1. Timo Z. Nazari-Shafti
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    2. Ivone G. Bruno
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    3. Rudy F. Martinez
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    4. Michael E. Coleman
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    5. Eckhard U. Alt
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    6. Scott R. McClure
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    Correspondence to Scott R. McClure .

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    Editors and Affiliations

    1. HemoGenix, Inc., Colorado Springs, Colorado, USA

      Ivan N. Rich

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    © 2015 Springer Science+Business Media New York

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    Nazari-Shafti, T.Z., Bruno, I.G., Martinez, R.F., Coleman, M.E., Alt, E.U., McClure, S.R. (2015). High Yield Recovery of Equine Mesenchymal Stem Cells from Umbilical Cord Matrix/Wharton’s Jelly Using a Semi-automated Process. In: Rich, I. (eds) Stem Cell Protocols. Methods in Molecular Biology, vol 1235. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1785-3_12

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    • DOI: https://doi.org/10.1007/978-1-4939-1785-3_12

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-1784-6

    • Online ISBN: 978-1-4939-1785-3

    • eBook Packages: Springer Protocols

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