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Exogenous expression of homeoprotein EGAM1N prevents in vitro cardiomyogenesis by impairing expression of T and Nkx2.5, but not Mef2c, in mouse embryonic stem cells

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Abstract

Generation of multiple cell types from embryonic stem (ES) cells and induced pluripotent stem cells is crucial to provide materials for regenerative medicine. EGAM1N has been found in preimplantation mouse embryos and mouse ES cells as a functionally unclassified homeoprotein. Recently, we reported that expression of EGAM1N suppressed the in vitro differentiation of ES cells into progenitor cells that arise in early embryogenesis. To clarify the effect of EGAM1N on terminal differentiation, embryoid bodies (EBs) were prepared from ES cells expressing exogenous Egam1n. In EBs expressing Egam1n, cardiomyogenesis was inhibited by impairing the expression of crucial transcription factors Brachyury T and Nkx2.5 in the generation of mesoderm and cardiomyocytes, respectively. Expression levels of Mef2c, another crucial gene for cardiomyogenesis, were unaffected. Conversely, the expression levels of Gata6 and Plat, markers for the primitive endoderm lineage, and Cdx2, a marker for the trophectoderm lineage, were increased. These results suggested that certain cell populations in EBs expressing Egam1n preferentially differentiated to such cell lineages. Our results suggest that EGAM1N not only affects the generation of progenitor cells during early embryogenesis, but also the progression of terminal differentiation, such as cardiomyogenesis, in mouse ES cells.

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Acknowledgments

We thank Dr. Max Gassmann (University of Zurich, Zurich, Switzerland) for providing the pMGD20neo vector and MG1.19 cells.

Funding

This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS KAKENHI No. 24580413) and the Akita Prefectural University President’s Research Project to M.K.

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    Authors and Affiliations

    1. Laboratory for Advanced Animal Cell Technology, Graduate School of Bioresource Sciences, Akita Prefectural University, Akita, 010-0195, Japan

      Asumi Nonaka, Michiko Yoshida, Momoe Iha, Yusuke Kubo, Yumi Kihara, Takahiro Kikuchi, Yuki Kumagai, Akira Sasaki & Masayuki Kobayashi

    2. Akita Research Institute of Food and Brewing, Akita, 010-1623, Japan

      Akira Sasaki

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    1. Asumi Nonaka
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    2. Michiko Yoshida
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    3. Momoe Iha
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    Correspondence to Masayuki Kobayashi.

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    A. Nonaka and M. Yoshida have contributed equally to this work.

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    Nonaka, A., Yoshida, M., Iha, M. et al. Exogenous expression of homeoprotein EGAM1N prevents in vitro cardiomyogenesis by impairing expression of T and Nkx2.5, but not Mef2c, in mouse embryonic stem cells. Cytotechnology 68, 2431–2436 (2016). https://doi.org/10.1007/s10616-016-9961-6

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