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Clump formation in mouse pituitary-derived non-endocrine cell line Tpit/F1 promotes differentiation into growth-hormone-producing cells

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Abstract

The adenohypophysis comprises six types of endocrine cells, including PIT1-lineage cells such as growth hormone (GH)-producing cells and heterogeneous non-endocrine cells, such as pituitary stem/progenitor cells as a source of endocrine cells. We determine the expression of characteristic stem cell marker genes, including sex-determining region Y-box 2 (Sox2), in mouse pituitary-derived non-endocrine cell lines Tpit/E, Tpit/F1 and TtT/GF. We observed high expression of fibroblast growth factor (FGF) receptors in Tpit/F1 cells, which we characterised by cultivation in medium containing a basic FGF and B27 supplement as used for neural stem-cell differentiation. A 4-day cultivation of Tpit/F1 produced floating embryonic stem-cell-like clumps accompanied by a three-fold increase in Sox2 expression. Passages in these clumps maintained the proliferative activity and Sox2 expression levels. After 10 days of cultivation, Tpit/F1 cell clumps were immuno-positive for SOX2 and Ki67 (proliferation marker) and loosely attached to the well bottom. An additional 10 days of cultivation induced the emergence of GH-positive/pituitary-specific transcription factor (PIT1)-negative cells showing migration from the clumps. Pit1 overexpression in attached cells could not induce GH production. Finally, we confirmed the presence of PIT1-negative GH-producing cells (3.2–7.7 % of all GH-positive cells) in rat pituitary. Thus, we demonstrate that Tpit/F1 has the plasticity to differentiate into one type of hormone-producing cell.

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Acknowledgements

This work was partially supported by JSPS KAKENHI grant numbers 15K18801 to M.H., 21380184 to Y.K. and 24580435 to T.K., by a MEXT-Supported Program for the Strategic Research Foundation at Private Universities 2014-2018 and by a research grant (A) to Y.K. from the Institute of Science and Technology, Meiji University. This study was also supported by the Meiji University International Institute for BioResource Research (MUIIR).

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  1. Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan

    Masashi Higuchi, Saishu Yoshida, Mo Chen, Takako Kato & Yukio Kato

  2. Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan

    Naoko Kanno, Hideo Mitsuishi, Hiroki Ueharu & Naoto Nishimura

  3. Department of Life Science, School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan

    Yukio Kato

  4. Joint Department of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori, 680-8553, Japan

    Masashi Higuchi

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  1. Masashi Higuchi
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Higuchi, M., Yoshida, S., Kanno, N. et al. Clump formation in mouse pituitary-derived non-endocrine cell line Tpit/F1 promotes differentiation into growth-hormone-producing cells. Cell Tissue Res 369, 353–368 (2017). https://doi.org/10.1007/s00441-017-2603-2

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