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      Human Mesenchymal Cells from Adipose Tissue Deposit Laminin and Promote Regeneration of Injured Spinal Cord in Rats

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          Abstract

          Cell therapy is a promising strategy to pursue the unmet need for treatment of spinal cord injury (SCI). Although several studies have shown that adult mesenchymal cells contribute to improve the outcomes of SCI, a descripton of the pro-regenerative events triggered by these cells is still lacking. Here we investigated the regenerative properties of human adipose tissue derived stromal cells (hADSCs) in a rat model of spinal cord compression. Cells were delivered directly into the spinal parenchyma immediately after injury. Human ADSCs promoted functional recovery, tissue preservation, and axonal regeneration. Analysis of the cord tissue showed an abundant deposition of laminin of human origin at the lesion site and spinal midline; the appearance of cell clusters composed of neural precursors in the areas of laminin deposition, and the appearance of blood vessels with separated basement membranes along the spinal axis. These effects were also observed after injection of hADSCs into non-injured spinal cord. Considering that laminin is a well-known inducer of axonal growth, as well a component of the extracellular matrix associated to neural progenitors, we propose that it can be the paracrine factor mediating the pro-regenerative effects of hADSCs in spinal cord injury.

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          Adult SVZ stem cells lie in a vascular niche: a quantitative analysis of niche cell-cell interactions.

          Qin Shen, Yue Wang, Erzsebet Kokovay … (2008)
          There is an emerging understanding of the importance of the vascular system within stem cell niches. Here, we examine whether neural stem cells (NSCs) in the adult subventricular zone (SVZ) lie close to blood vessels, using three-dimensional whole mounts, confocal microscopy, and automated computer-based image quantification. We found that the SVZ contains a rich plexus of blood vessels that snake along and within neuroblast chains. Cells expressing stem cell markers, including GFAP, and proliferation markers are closely apposed to the laminin-containing extracellular matrix (ECM) surrounding vascular endothelial cells. Apical GFAP+ cells are admixed within the ependymal layer and some span between the ventricle and blood vessels, occupying a specialized microenvironment. Adult SVZ progenitor cells express the laminin receptor alpha6beta1 integrin, and blocking this inhibits their adhesion to endothelial cells, altering their position and proliferation in vivo, indicating that it plays a functional role in binding SVZ stem cells within the vascular niche.
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            Mesenchymal stem cell-educated macrophages: a novel type of alternatively activated macrophages.

            Jaehyup Kim, Peiman Hematti (2009)
            Mesenchymal stem cells (MSCs) are capable of modulating the immune system through interaction with a wide range of immune cells. This study investigates the hypothesis that interaction of MSCs with macrophages could play a significant role in their antiinflammatory/immune modulatory effects. MSCs were derived from bone marrow and monocytes were isolated from peripheral blood of healthy donors. We cultured human monocytes for 7 days without any added cytokines to generate macrophages, and then cocultured them for 3 more days with culture-expanded MSCs. We used cell surface antigen expression and intracellular cytokine expression patterns to study the immunophenotype of macrophages at the end of this coculture period, and phagocytic assays to investigate their functional activity in vitro. Macrophages cocultured with MSCs consistently showed high-level expression of CD206, a marker of alternatively activated macrophages. Furthermore, these macrophages expressed high levels of interleukin (IL)-10 and low levels of IL-12, as determined by intracellular staining, typical of alternatively activated macrophages. However, macrophages cocultured with MSCs also expressed high levels of IL-6 and low levels of tumor necrosis factor-alpha (TNF-alpha) compared to controls. Functionally, macrophages cocultured with MSCs showed a higher level of phagocytic activity. We describe a novel type of human macrophage generated in vitro after coculture with MSCs that assumes an immunophenotype defined as IL-10-high, IL-12-low, IL-6-high, and TNF-alpha-low secreting cells. These MSC-educated macrophages may be a unique and novel type of alternatively activated macrophage with a potentially significant role in tissue repair.
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              GAP-43: an intrinsic determinant of neuronal development and plasticity.

              Larry I. Benowitz, Aryeh Routtenberg (1997)
              Several lines of investigation have helped clarify the role of GAP-43 (FI, B-50 or neuromodulin) in regulating the growth state of axon terminals. In transgenic mice, overexpression of GAP-43 leads to the spontaneous formation of new synapses and enhanced sprouting after injury. Null mutation of the GAP-43 gene disrupts axonal pathfinding and is generally lethal shortly after birth. Manipulations of GAP-43 expression likewise have profound effects on neurite outgrowth for cells in culture. GAP-43 appears to be involved in transducing intra- and extracellular signals to regulate cytoskeletal organization in the nerve ending. Phosphorylation by protein kinase C is particularly significant in this regard, and is linked with both nerve-terminal sprouting and long-term potentiation. In the brains of humans and other primates, high levels of GAP-43 persist in neocortical association areas and in the limbic system throughout life, where the protein might play an important role in mediating experience-dependent plasticity.
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                Author and article information

                Contributors
                Eva Mezey: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, USA )
                ISSN (Electronic): 1932-6203
                Publication date Collection: 2014
                Publication date (Electronic): 15 May 2014
                Volume: 9
                Issue: 5
                Electronic Location Identifier: e96020
                Affiliations
                [1 ]Institute of Biomedical Sciences, Federal University of Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil
                [2 ]Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Rio de Janeiro, Brazil
                [3 ]National Institute of Cancer, Rio de Janeiro, Rio de Janeiro, Brazil
                [4 ]Excellion, Petrópolis, Rio de Janeiro, Brazil
                National Institutes of Health, United States of America
                Author notes

                Competing Interests: Radovan Borojevic is an employee of the company Excellion. This does not alter the authors’ adherence to all the PLOS ONE policies on sharing data and materials.

                Conceived and designed the experiments: KM MB JRLM RB MIDR TCS. Performed the experiments: KM MAN JPG ASC DVL BC JRLM. Analyzed the data: KM MAN JRLM RB MIDR TCS. Contributed reagents/materials/analysis tools: MB JRLM RB MIDR TCS. Wrote the paper: KM JRLM MIDR TCS.

                Article
                Publisher ID: PONE-D-13-08621
                DOI: 10.1371/journal.pone.0096020
                PMC ID: 4022508
                PubMed ID: 24830794
                SO-VID: 800c29cf-ccc1-40d8-a05a-061f4a5bdb3f
                Copyright statement: Copyright @ 2014
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                Date received : 25 February 2013
                Date accepted : 3 April 2014
                Page count
                Pages: 15
                Funding
                This work was supported by grants from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Fundação Carlos Chagas Filho de Apoio à Pesquisa do Estado do Rio de Janeiro (FAPERJ). KM and JPG are recipients of fellowships from CNPq, and MAN and DVL received fellowships from Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior (CAPES) and FAPERJ, respectively. ASC received an undergraduate fellowship from CNPq. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Subject: Research Article
                Subject: Biology and Life Sciences
                Subject: Anatomy
                Subject: Nervous System
                Subject: Cell Biology
                Subject: Cellular Structures and Organelles
                Subject: Extracellular Matrix
                Subject: Cellular Types
                Subject: Animal Cells
                Subject: Stem Cells
                Subject: Mesenchymal Stem Cells
                Subject: Molecular Cell Biology
                Subject: Developmental Biology
                Subject: Neuroscience
                Subject: Cellular Neuroscience
                Subject: Organisms
                Subject: Animals
                Subject: Vertebrates
                Subject: Mammals
                Subject: Rodents
                Subject: Rats
                Subject: Medicine and Health Sciences
                Subject: Neurology
                Subject: Neurobiology of Disease and Regeneration
                Subject: Research and Analysis Methods
                Subject: Model Organisms
                Subject: Animal Models

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