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      Isolation and characterization of GFAP-positive porcine neural stem/progenitor cells derived from a GFAP-CreER T2 transgenic piglet

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          Abstract

          Background

          The porcine brain is gyrencephalic with similar gray and white matter composition and size more comparable to the human rather than the rodent brain; however, there is lack of information about neural progenitor cells derived from this model.

          Results

          Here, we isolated GFAP-positive porcine neural stem cells (NSCs) from the brain explant of a transgenic piglet, with expression of CreER T2 under the control of the GFAP promoter (pGFAP-CreER T2). The isolated pGFAP-CreER T2 NSCs showed self-renewal and expression of representative NSC markers such as Nestin and Sox2. Pharmacological inhibition studies revealed that Notch1 signaling is necessary to maintain NSC identity, whereas serum treatment induced cell differentiation into reactive astrocytes and neurons.

          Conclusions

          Collectively, these results indicate that GFAP promoter-driven porcine CreER T2 NSCs would be a useful tool to study neurogenesis of the porcine adult central nervous system and furthers our understanding of its potential clinical application in the future.

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          Electronic supplementary material

          The online version of this article (10.1186/s12917-018-1660-4) contains supplementary material, which is available to authorized users.

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          Most cited references38

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          Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system.

          Neurogenesis in the mammalian central nervous system is believed to end in the period just after birth; in the mouse striatum no new neurons are produced after the first few days after birth. In this study, cells isolated from the striatum of the adult mouse brain were induced to proliferate in vitro by epidermal growth factor. The proliferating cells initially expressed nestin, an intermediate filament found in neuroepithelial stem cells, and subsequently developed the morphology and antigenic properties of neurons and astrocytes. Newly generated cells with neuronal morphology were immunoreactive for gamma-aminobutyric acid and substance P, two neurotransmitters of the adult striatum in vivo. Thus, cells of the adult mouse striatum have the capacity to divide and differentiate into neurons and astrocytes.
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            Coexistence of quiescent and active adult stem cells in mammals.

            Adult stem cells are crucial for physiological tissue renewal and regeneration after injury. Prevailing models assume the existence of a single quiescent population of stem cells residing in a specialized niche of a given tissue. Emerging evidence indicates that both quiescent (out of cell cycle and in a lower metabolic state) and active (in cell cycle and not able to retain DNA labels) stem cell subpopulations may coexist in several tissues, in separate yet adjoining locations. Here, we summarize these findings and propose that quiescent and active stem cell populations have separate but cooperative functional roles.
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              Neural stem cells confer unique pinwheel architecture to the ventricular surface in neurogenic regions of the adult brain.

              Neural stem cells (NSCs, B1 cells) are retained in the walls of the adult lateral ventricles but, unlike embryonic NSCs, are displaced from the ventricular zone (VZ) into the subventricular zone (SVZ) by ependymal cells. Apical and basal compartments, which in embryonic NSCs play essential roles in self-renewal and differentiation, are not evident in adult NSCs. Here we show that SVZ B1 cells in adult mice extend a minute apical ending to directly contact the ventricle and a long basal process ending on blood vessels. A closer look at the ventricular surface reveals a striking pinwheel organization specific to regions of adult neurogenesis. The pinwheel's core contains the apical endings of B1 cells and in its periphery two types of ependymal cells: multiciliated (E1) and a type (E2) characterized by only two cilia and extraordinarily complex basal bodies. These results reveal that adult NSCs retain fundamental epithelial properties, including apical and basal compartmentalization, significantly reshaping our understanding of this adult neurogenic niche.
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                Author and article information

                Contributors
                eunhye.kim2@emory.edu
                ghkdsun@hanmail.net
                padugey@gmail.com
                hg-kim@korea.ac.kr
                glee48@jhmi.edu
                +82-43-261-3393 , shhyun@cbu.ac.kr
                Journal
                BMC Vet Res
                BMC Vet. Res
                BMC Veterinary Research
                BioMed Central (London )
                1746-6148
                7 November 2018
                7 November 2018
                2018
                : 14
                : 331
                Affiliations
                [1 ]ISNI 0000 0000 9611 0917, GRID grid.254229.a, Laboratory of Veterinary Embryology and Biotechnology, Veterinary Medical Center and College of Veterinary Medicine, , Chungbuk National University, ; 1 Chungdae-ro, Seowon-gu, Cheongju, 28644 Republic of Korea
                [2 ]ISNI 0000 0000 9611 0917, GRID grid.254229.a, Institute of Stem Cell & Regenerative Medicine (ISCRM), , Chungbuk National University, ; Cheongju, 28644 Chungbuk Republic of Korea
                [3 ]ISNI 0000 0001 0840 2678, GRID grid.222754.4, Department of Biotechnology, School of Life Sciences and Biotechnology, , Korea University, ; 02841 Seoul, Republic of Korea
                [4 ]ISNI 0000 0001 2171 9311, GRID grid.21107.35, Institute for Cell Engineering, , Johns Hopkins University School of Medicine, ; Baltimore, MD USA
                Author information
                http://orcid.org/0000-0001-6011-2555
                Article
                1660
                10.1186/s12917-018-1660-4
                6222979
                30404643
                6caae641-7ffe-4058-929c-81f985277297
                © The Author(s). 2018

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 19 May 2018
                : 22 October 2018
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100003725, National Research Foundation of Korea;
                Award ID: NRF-2016R1D1A1B03933191, NRF-2017R1A2B4002546, 2017K1A4A3014959
                Funded by: Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET)
                Award ID: 115103-02
                Funded by: Business for Cooperative R&D between Industry, Academy, and Research Institute
                Award ID: 2017020681010101
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2018

                Veterinary medicine
                pig,neural stem cells,notch signaling,reactive astrocytes
                Veterinary medicine
                pig, neural stem cells, notch signaling, reactive astrocytes

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