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      Harnessing a Feasible and Versatile ex vivo Calvarial Suture 2-D Culture System to Study Suture Biology

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          Abstract

          As a basic science, craniofacial research embraces multiple facets spanning from molecular regulation of craniofacial development, cell biology/signaling and ultimately translational craniofacial biology. Calvarial sutures coordinate development of the skull, and the premature fusion of one or more, leads to craniosynostosis. Animal models provide significant contributions toward craniofacial biology and clinical/surgical treatments of patients with craniofacial disorders. Studies employing mouse models are costly and time consuming for housing/breeding. Herein, we present the establishment of a calvarial suture explant 2-D culture method that has been proven to be a reliable system showing fidelity with the in vivo harvesting procedure to isolate high yields of skeletal stem/progenitor cells from small number of mice. Moreover, this method allows the opportunity to phenocopying models of craniosynostosis and in vitro tamoxifen-induction of Actin creERT2;R26 Rainbow suture explants to trace clonal expansion. This versatile method tackles needs of large number of mice to perform calvarial suture research.

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          Most cited references52

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          The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2018 update

          Abstract Galaxy (homepage: https://galaxyproject.org, main public server: https://usegalaxy.org) is a web-based scientific analysis platform used by tens of thousands of scientists across the world to analyze large biomedical datasets such as those found in genomics, proteomics, metabolomics and imaging. Started in 2005, Galaxy continues to focus on three key challenges of data-driven biomedical science: making analyses accessible to all researchers, ensuring analyses are completely reproducible, and making it simple to communicate analyses so that they can be reused and extended. During the last two years, the Galaxy team and the open-source community around Galaxy have made substantial improvements to Galaxy's core framework, user interface, tools, and training materials. Framework and user interface improvements now enable Galaxy to be used for analyzing tens of thousands of datasets, and >5500 tools are now available from the Galaxy ToolShed. The Galaxy community has led an effort to create numerous high-quality tutorials focused on common types of genomic analyses. The Galaxy developer and user communities continue to grow and be integral to Galaxy's development. The number of Galaxy public servers, developers contributing to the Galaxy framework and its tools, and users of the main Galaxy server have all increased substantially.
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            The stem-cell niche as an entity of action.

            Stem-cell populations are established in 'niches'--specific anatomic locations that regulate how they participate in tissue generation, maintenance and repair. The niche saves stem cells from depletion, while protecting the host from over-exuberant stem-cell proliferation. It constitutes a basic unit of tissue physiology, integrating signals that mediate the balanced response of stem cells to the needs of organisms. Yet the niche may also induce pathologies by imposing aberrant function on stem cells or other targets. The interplay between stem cells and their niche creates the dynamic system necessary for sustaining tissues, and for the ultimate design of stem-cell therapeutics.
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              Krüppel-like factor 4 (KLF4): What we currently know.

              Krüppel-like factor 4 (KLF4) is an evolutionarily conserved zinc finger-containing transcription factor that regulates diverse cellular processes such as cell growth, proliferation, and differentiation. Since its discovery in 1996, KLF4 has been gaining a lot of attention, particularly after it was shown in 2006 as one of four factors involved in the induction of pluripotent stem cells (iPSCs). Here we review the current knowledge about the different functions and roles of KLF4 in various tissue and organ systems.
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                Author and article information

                Contributors
                Journal
                Front Physiol
                Front Physiol
                Front. Physiol.
                Frontiers in Physiology
                Frontiers Media S.A.
                1664-042X
                10 February 2022
                2022
                : 13
                : 823661
                Affiliations
                [1] 1Hagey Laboratory for Pediatric Regenerative Medicine, Stanford University School of Medicine , Stanford, CA, United States
                [2] 2Division of Plastic and Reconstructive Surgery, Stanford University School of Medicine , Stanford, CA, United States
                [3] 3Department of Surgery, Stanford University School of Medicine , Stanford, CA, United States
                [4] 4Dipartimento di Scienze Biomediche Avanzate, Università degli Studi di Napoli Federico II , Naples, Italy
                [5] 5Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine , Stanford, CA, United States
                [6] 6Department of Plastic Surgery, BG University Hospital Bergmannsheil Bochum , Bochum, Germany
                Author notes

                Edited by: Timothy C. Cox, University of Missouri–Kansas City, United States

                Reviewed by: Nan E. Hatch, University of Michigan, United States; Jacqueline Tabler, Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society, Germany

                *Correspondence: Natalina Quarto, nquarto@ 123456stanford.edu
                Michael T. Longaker, longaker@ 123456stanford.edu

                These authors have contributed equally to this work

                This article was submitted to Craniofacial Biology and Dental Research, a section of the journal Frontiers in Physiology

                Article
                10.3389/fphys.2022.823661
                8871685
                de2a4636-9c61-4826-b1fd-b08a265a9a5f
                Copyright © 2022 Quarto, Menon, Griffin, Huber and Longaker.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 27 November 2021
                : 13 January 2022
                Page count
                Figures: 7, Tables: 0, Equations: 0, References: 52, Pages: 13, Words: 8419
                Funding
                Funded by: National Institutes of Health, doi 10.13039/100000002;
                Award ID: R01DE027323, R01DE026730, U24DE029463
                Categories
                Physiology
                Methods

                Anatomy & Physiology
                calvarial suture,ex vivo explants,culture method,biology,phenocopy craniosynostosis

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