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      Organ In Vitro Culture: What Have We Learned about Early Kidney Development?

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          Abstract

          When Clifford Grobstein set out to study the inductive interaction between tissues in the developing embryo, he developed a method that remained important for the study of renal development until now. From the late 1950s on, in vitro cultivation of the metanephric kidney became a standard method. It provided an artificial environment that served as an open platform to study organogenesis. This review provides an introduction to the technique of organ culture, describes how the Grobstein assay and its variants have been used to study aspects of mesenchymal induction, and describes the search for natural and chemical inducers of the metanephric mesenchyme. The review also focuses on renal development, starting with ectopic budding of the ureteric bud, ureteric bud branching, and the generation of the nephron and presents the search for stem cells and renal progenitor cells that contribute to specific structures and tissues during renal development. It also presents the current use of Grobstein assay and its modifications in regenerative medicine and tissue engineering today. Together, this review highlights the importance of ex vivo kidney studies as a way to acquire new knowledge, which in the future can and will be implemented for developmental biology and regenerative medicine applications.

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          Establishment in culture of pluripotential cells from mouse embryos.

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            A series of normal stages in the development of the chick embryo

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              Six2 defines and regulates a multipotent self-renewing nephron progenitor population throughout mammalian kidney development.

              Nephrons, the basic functional units of the kidney, are generated repetitively during kidney organogenesis from a mesenchymal progenitor population. Which cells within this pool give rise to nephrons and how multiple nephron lineages form during this protracted developmental process are unclear. We demonstrate that the Six2-expressing cap mesenchyme represents a multipotent nephron progenitor population. Six2-expressing cells give rise to all cell types of the main body of the nephron during all stages of nephrogenesis. Pulse labeling of Six2-expressing nephron progenitors at the onset of kidney development suggests that the Six2-expressing population is maintained by self-renewal. Clonal analysis indicates that at least some Six2-expressing cells are multipotent, contributing to multiple domains of the nephron. Furthermore, Six2 functions cell autonomously to maintain a progenitor cell status, as cap mesenchyme cells lacking Six2 activity contribute to ectopic nephron tubules, a mechanism dependent on a Wnt9b inductive signal. Taken together, our observations suggest that Six2 activity cell-autonomously regulates a multipotent nephron progenitor population.
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                Author and article information

                Journal
                Stem Cells Int
                Stem Cells Int
                SCI
                Stem Cells International
                Hindawi Publishing Corporation
                1687-966X
                1687-9678
                2015
                19 May 2015
                : 2015
                : 959807
                Affiliations
                1Faculty of Biochemistry and Molecular Medicine, Biocenter Oulu, Oulu University, 90220 Oulu, Finland
                2Renal Regeneration Laboratory, VAGLAHS at Sepulveda, North Hills, CA 91343, USA
                3David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA
                Author notes
                *Aleksandra Rak-Raszewska: aleksandra.rak-raszewska@ 123456oulu.fi and
                *Peter V. Hauser: pvhauser@ 123456ucla.edu

                Academic Editor: Laura Lasagni

                Article
                10.1155/2015/959807
                4452498
                26078765
                a5ef5efe-79ae-4c67-8e3c-88cb1a8f32ba
                Copyright © 2015 Aleksandra Rak-Raszewska et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 7 November 2014
                : 7 January 2015
                : 8 January 2015
                Categories
                Review Article

                Molecular medicine
                Molecular medicine

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