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      Feeder Cells Support the Culture of Induced Pluripotent Stem Cells Even after Chemical Fixation

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          Abstract

          Chemically fixed mouse embryonic fibroblasts (MEFs), instead of live feeder cells, were applied to the maintenance of mouse induced pluripotent stem (miPS) cells. Formaldehyde and glutaraldehyde were used for chemical fixation. The chemically fixed MEF feeders maintained the pluripotency of miPS cells, as well as their undifferentiated state. Furthermore, the chemically fixed MEF feeders were reused several times without affecting their functions. These results indicate that chemical fixation can be applied to modify biological feeders chemically, without losing their original functions. Chemically fixed MEF feeders will be applicable to other stem cell cultures as a reusable extracellular matrix candidate that can be preserved on a long-term basis.

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          Most cited references22

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          Establishment in culture of pluripotential cells from mouse embryos.

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            Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells.

            Embryonic stem (ES) cells, the totipotent outgrowths of blastocysts, can be cultured and manipulated in vitro and then returned to the embryonic environment where they develop normally and can contribute to all cell lineages. Maintenance of the stem-cell phenotype in vitro requires the presence of a feeder layer of fibroblasts or of a soluble factor, differentiation inhibitory activity (DIA) produced by a number of sources; in the absence of DIA the ES cells differentiate into a wide variety of cell types. We recently noted several similarities between partially purified DIA and a haemopoietic regulator, myeloid leukaemia inhibitory factor (LIF), a molecule which induces differentiation in M1 myeloid leukaemic cells and which we have recently purified, cloned and characterized. We demonstrate here that purified, recombinant LIF can substitute for DIA in the maintenance of totipotent ES cell lines that retain the potential to form chimaeric mice.
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              Derivation of human embryonic stem cells in defined conditions.

              We have previously reported that high concentrations of basic fibroblast growth factor (bFGF) support feeder-independent growth of human embryonic stem (ES) cells, but those conditions included poorly defined serum and matrix components. Here we report feeder-independent human ES cell culture that includes protein components solely derived from recombinant sources or purified from human material. We describe the derivation of two new human ES cell lines in these defined culture conditions.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                2 March 2012
                : 7
                : 3
                : e32707
                Affiliations
                [1 ]Nano Medical Engineering Laboratory, RIKEN Advanced Science Institute, Wako-shi, Saitama, Japan
                [2 ]Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama-shi, Kanagawa, Japan
                [3 ]Support Unit for Animal Resources Development, Research Resources Center, RIKEN Brain Science Institute, Wako-shi, Saitama, Japan
                Baylor College of Medicine, United States of America
                Author notes

                Conceived and designed the experiments: YI. Performed the experiments: X-SY MF CN J-SG T. Arai. Analyzed the data: X-SY ET YI. Contributed reagents/materials/analysis tools: MF CN T. Akaike ET. Wrote the paper: X-SY YI.

                [¤]

                Current address: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana, United States of America

                Article
                PONE-D-11-19849
                10.1371/journal.pone.0032707
                3292564
                22396791
                3c9d63a4-b32a-4a3c-b52d-c053e4614925
                Yue et al. 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
                : 10 October 2011
                : 1 February 2012
                Page count
                Pages: 9
                Categories
                Research Article
                Biology
                Biotechnology
                Developmental Biology
                Stem Cells
                Molecular Cell Biology
                Cellular Types
                Stem Cells
                Materials Science
                Biomaterials

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