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          Institute: MPI für molekulare Biomedizin     Collection: Jahrbuch 2017 (publ. 2016, arch)     Display Documents



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ID: 732142.0, MPI für molekulare Biomedizin / Jahrbuch 2017 (publ. 2016, arch)
Biphasic modulation of Wnt signaling supports efficient foregut endoderm formation from human pluripotent stem cells
Authors:Hoepfner, J.; Kleinsorge, M.; Papp, O.; Ackermann, M.; Alfken, S.; Rinas, U.; Solodenko, W.; Kirschning, A.; Sgodda, M.; Cantz, T.
Date of Publication (YYYY-MM-DD):2016-05
Title of Journal:Cell Biol Int
Volume:40
Issue / Number:5
Start Page:534
End Page:548
Review Status:Internal review
Audience:Not Specified
Abstract / Description:Pluripotent stem cells (embryonic stem cells and induced pluripotent stem cells) are of great promise in regenerative medicine, including molecular studies of disease mechanisms, if the affected cell type can be authentically generated during in vitro differentiation. Most existing protocols aim to mimic embryonic development steps by the supplementation of specific cytokines and small molecules, but the involved signaling pathways need further exploration. In this study, we investigated enhanced initial activation of Wnt signaling for definitive endoderm formation and subsequent rapid shutdown of Wnt signaling for proper foregut endoderm specification using 3 muM CHIR99021 and 0.5 mug/mL of secreted frizzled-related protein 5 (sFRP-5) for biphasic modulation of the Wnt pathway. The definitive endoderm and foregut endoderm differentiation capabilities of Wnt pathway-modulated cells were determined based on the expression levels of the endodermal transcription factors SOX17 and FOXA2 and those of the transcription activator GATA4 and the alpha-fetoprotein (AFP) gene, respectively. Furthermore, the resulting biphasic Wnt pathway modulation was investigated at the protein level by analyzing phosphorylation of glycogen synthase kinase 3 beta (GSK3beta) and beta-catenin. Finally, Wnt target gene expression was determined using an improved lentiviral reporter construct that enabled robust T-cell transcription factor 4 (TCF4)/lymphoid enhancer-binding factor 1 (LEF1)-mediated luciferase expression in differentiating pluripotent stem cells. In conclusion, we demonstrated robust, homogeneous, and efficient derivation of foregut endodermal cells by inducing a biphasic modulation of the Wnt signaling pathway.
Free Keywords:cell differentiation; liver/hepatocytes; stem cells
External Publication Status:published
Document Type:Article
Communicated by:Jeanine Müller-Keuker
Affiliations:MPI für molekulare Biomedizin
External Affiliations:Department of Gastroenterology, Hepatology, and Endocrinology, Hannover Medical School, Hannover, Germany. iPSC Based Gene Therapy, REBIRTH Cluster of Excellence, Hannover Medical School, Hannover, Germany. Institute of Technical Chemistry, Leibniz University Hannover, Hannover, Germany. Institute of Organic Chemistry, Leibniz University Hannover, Hannover, Germany. Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Munster, Germany.
Identifiers:ISSN:1095-8355 (Electronic) 1065-6995 (Linking) %R 10.1... [ID No:1]
URL:https://www.ncbi.nlm.nih.gov/pubmed/26861571 [ID No:2]
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