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      Comparison between adult and foetal adnexa derived equine post-natal mesenchymal stem cells

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          Abstract

          Background

          Little is known about the differences among adult and foetal equine mesenchymal stem cells (MSCs), and no data exist about their comparative ultrastructural morphology. The aim of this study was to describe and compare characteristics, immune properties, and ultrastructural morphology of equine adult (bone marrow: BM, and adipose tissue: AT) and foetal adnexa derived (umbilical cord blood: UCB, and Wharton’s jelly: WJ) MSCs.

          Results

          No differences were observed in proliferation during the first 3 passages. While migration ability was similar among cells, foetal MSCs showed a higher adhesion ability, forming smaller spheroids after hanging drop culture ( P < 0.05). All MSCs differentiated toward adipogenic, chondrogenic and osteogenic lineages, only tenogenic differentiation was less evident for WJ-MSCs. Data obtained by PCR confirmed MHC1 expression and lack of MHC2 expression in all four cell types. Foetal adnexa MSCs were positive for genes specific for anti-inflammatory and angiogenic factors (IL6, IL8, ILβ1) and WJ-MSCs were the only positive for OCT4 pluripotency gene. At immunofluorescence all cells expressed typical mesenchymal markers (α-SMA, N-cadherin), except for BM-MSCs, which did not express N-cadherin. By transmission electron microscopy, it was observed that WJ-MSCs had a higher ( P < 0.05) number of microvesicles compared to adult MSCs, and UCB-MSCs showed more microvesicles than BM-MSCs ( P < 0.05). AT-MSCs had a lower number of mitochondria than WJ-MSCs ( P < 0.05), and mitochondrial area was higher for WJ-MSCs compared to UCB and AT-MSCs ( P < 0.05).

          Conclusions

          Results demonstrate that MSCs from adult and foetal tissues have different characteristics, and foetal MSCs, particularly WJ derived ones, seem to have some charactestics that warrant further investigation into potential advantages for clinical application.

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          Most cited references46

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          Endothelial cell migration during angiogenesis.

          Endothelial cell migration is essential to angiogenesis. This motile process is directionally regulated by chemotactic, haptotactic, and mechanotactic stimuli and further involves degradation of the extracellular matrix to enable progression of the migrating cells. It requires the activation of several signaling pathways that converge on cytoskeletal remodeling. Then, it follows a series of events in which the endothelial cells extend, contract, and throw their rear toward the front and progress forward. The aim of this review is to give an integrative view of the signaling mechanisms that govern endothelial cell migration in the context of angiogenesis.
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            Three-dimensional aggregates of mesenchymal stem cells: cellular mechanisms, biological properties, and applications.

            Mesenchymal stem cells (MSCs) are primary candidates in cell therapy and tissue engineering and are being tested in clinical trials for a wide range of diseases. Originally isolated and expanded as plastic adherent cells, MSCs have intriguing properties of in vitro self-assembly into three-dimensional (3D) aggregates reminiscent of skeletal condensation in vivo. Recent studies have shown that MSC 3D aggregation improved a range of biological properties, including multilineage potential, secretion of therapeutic factors, and resistance against ischemic condition. Hence, the formation of 3D MSC aggregates has been explored as a novel strategy to improve cell delivery, functional activation, and in vivo retention to enhance therapeutic outcomes. This article summarizes recent reports of MSC aggregate self-assembly, characterization of biological properties, and their applications in preclinical models. The cellular and molecular mechanisms underlying MSC aggregate formation and functional activation are discussed, and the areas that warrant further investigation are highlighted. These analyses are combined to provide perspectives for identifying the controlling mechanisms and refining the methods of aggregate fabrication and expansion for clinical applications.
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              3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells.

              Multipotent mesenchymal stem cells (MSCs) are one of the most powerful tools in regeneration medicine. Their low differentiation efficiency, however, limits further application of MSCs in clinical therapy. Here we report that a much higher multipotent differentiation efficiency of MSCs to adult cells can be achieved using a 3D spheroid culture method based on photolithography and micropatterning techniques. MSC spheroid of precise dimension and uniform quality cultured on the microdomain substrates was prepared first, and then was induced into adipocytes and osteoblasts. Both gene expression results from RT-PCR and morphology observation results revealed that the 3D spheroid culture method could greatly improve differentiation efficiency. Gene expression profiles obtained from gene microarray analysis confirmed the high differentiation efficiency and revealed that MSCs induced in 3D spheroid culture system regulated gene expression not only by increasing the expression levels of genes related to adipogenesis and osteogenesis, but also by down-regulating the gene maintaining MSCs' self-renewal phenotypes. We conclude that our 3D spheroid culture system contributes to an optimization for efficient differentiation of MSCs, offers insight into the mechanism of efficient differentiation of engineered 3D culture system, and has promise for wide applications in regeneration medicine and drug discovery fields.
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                Author and article information

                Contributors
                barbara.merlo@unibo.it
                gabriella.teti2@unibo.it
                aliai.lanci2@unibo.it
                janina.burk@vetmed.uni-giessen.de
                emazzotti@unite.it
                mirella.falconi@unibo.it
                eleonora.iacono2@unibo.it
                Journal
                BMC Vet Res
                BMC Vet. Res
                BMC Veterinary Research
                BioMed Central (London )
                1746-6148
                2 August 2019
                2 August 2019
                2019
                : 15
                : 277
                Affiliations
                [1 ]ISNI 0000 0004 1757 1758, GRID grid.6292.f, Department of Veterinary Medical Sciences, , University of Bologna, ; via Tolara di Sopra 50, 40064 Ozzano Emilia, BO Italy
                [2 ]ISNI 0000 0004 1757 1758, GRID grid.6292.f, Department for Biomedical and Neuromotor Sciences, , University of Bologna, ; Bologna, Italy
                [3 ]ISNI 0000 0001 2230 9752, GRID grid.9647.c, Saxon Incubator for Clinical Translation, , University of Leipzig, ; Leipzig, Germany
                [4 ]ISNI 0000 0001 2165 8627, GRID grid.8664.c, Equine Clinic (Surgery), , Justus Liebig University Giessen, ; Giessen, Germany
                [5 ]ISNI 0000 0001 2202 794X, GRID grid.17083.3d, Department of Comparative Biomedical Sciences, , University of Teramo, ; Teramo, Italy
                [6 ]ISNI 0000 0004 1757 1758, GRID grid.6292.f, Health Science and Technologies Interdepartmental Center for Industrial Research (HST-ICIR), , University of Bologna, ; Bologna, Italy
                Author information
                http://orcid.org/0000-0002-4435-1844
                Article
                2023
                10.1186/s12917-019-2023-5
                6679462
                31375144
                58b30bca-a21e-4ec5-bbbd-e3301a880407
                © The Author(s). 2019

                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
                : 28 September 2018
                : 26 July 2019
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2019

                Veterinary medicine
                horse,mesenchymal stem cells,adult, foetal adnexa,transmission electron microscopy

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