title: Clump formation in mouse pituitary-derived non-endocrine cell line Tpit/F1 promotes differentiation into growth-hormone-producing cells.
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.
authors.affiliation: Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan.|Joint Department of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori, 680-8553, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Higuchi
authors.affiliation: Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Saishu
authors.initials: S
authors.lastname: Yoshida
authors.affiliation: Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Kanno
authors.affiliation: Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Mitsuishi
authors.affiliation: Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ueharu
authors.affiliation: Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Mo
authors.initials: M
authors.lastname: Chen
authors.affiliation: Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Nishimura
authors.affiliation: Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Kato
authors.affiliation: Institute for Reproduction and Endocrinology, Meiji University, Kanagawa, 214-8571, Japan. yukato@meiji.ac.jp.|Department of Life Science, School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan. yukato@meiji.ac.jp.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Kato
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion; D002449:Cell Aggregation; D002454:Cell Differentiation; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D048430:Cell Shape; D002864:Chromogranins / Q000378:metabolism; D003470:Culture Media; D055098:Endocrine Cells / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000096:biosynthesis*; D051379:Mice; D010902:Pituitary Gland / Q000166:cytology*; D051381:Rats; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D015415:Biomarkers; D002864:Chromogranins; D003470:Culture Media; D017468:Receptors, Fibroblast Growth Factor; D014157:Transcription Factors; D013006:Growth Hormone
keywords: Clump formation; Differentiation; Growth hormone; Stemness; Tpit/F1 cell