title: Effect of ectopic expression of homeoprotein EGAM1C on the cell morphology, growth, and differentiation in a mouse embryonic stem cell line, MG1.19 cells.
abstract: The homeoprotein EGAM1C was identified in preimplantation mouse embryos and embryonic stem (ES) cells. To explore the impact of EGAM1C on the hallmarks of mouse ES cells, MG1.19 cells stably expressing EGAM1C at levels similar to those in blastocysts were established using an episomal expression system. In the presence of leukemia inhibitory factor (+LIF), control transfectants with an empty vector formed flattened cell colonies, while Egam1c transfectants formed compacted colonies with increased E-CADHERIN expression. In Egam1c transfectants, the cellular contents of POU5F1 (OCT4), SOX2, TBX3, and NANOG increased. Cell growth was accelerated in an undifferentiated state sustained by LIF and in the course of differentiation. During clonal proliferation, EGAM1C stabilized the undifferentiated state. In adherent culture conditions, EGAM1C partly inhibited the progression of differentiation at least within a 4-day culture period in the presence of retinoic acid by preventing the downregulation of LIF signaling with a robust increase in TBX3 expression. Conversely, EGAM1C enhanced the expression of lineage marker genes Fgf5 (epiblast), T (mesoderm), Gata6 (primitive endoderm), and Cdx2 (trophectoderm) in -LIF conditions. In embryoid bodies expressing EGAM1C, the expression of marker genes for extraembryonic cell lineages, including Tpbpa (spongiotrophoblast) and Plat (parietal endoderm), increased. These results demonstrated that the ectopic expression of EGAM1C is capable of affecting the stabilization of an undifferentiated state and the progression of differentiation in MG1.19 ES cells, in addition to affecting cellular morphology and growth.
authors.affiliation: Laboratory for Advanced Animal Cell Technology, Graduate School of Bioresource Sciences, Akita Prefectural University, Akita 010-0195, Japan.
authors.forename: Momoe
authors.initials: M
authors.lastname: Iha
authors.forename: Masafumi
authors.initials: M
authors.lastname: Watanabe
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kihara
authors.forename: Saiko
authors.initials: S
authors.lastname: Sugawara
authors.forename: Koichi
authors.initials: K
authors.lastname: Saito
authors.forename: Miki
authors.initials: M
authors.lastname: Soma
authors.forename: Sho
authors.initials: S
authors.lastname: Sato
authors.forename: Yuki
authors.initials: Y
authors.lastname: Mori
authors.forename: Kano
authors.initials: K
authors.lastname: Kasuga
authors.forename: Ikuo
authors.initials: I
authors.lastname: Kojima
authors.forename: Ryo
authors.initials: R
authors.lastname: Sasamura
authors.forename: Jun
authors.initials: J
authors.lastname: Murata
authors.forename: Masayuki
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D058732:Embryoid Bodies / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D018398:Homeodomain Proteins / Q000378:metabolism*; D051379:Mice; D010957:Plasmids
chemical_list: C558397:EGAM1C protein, mouse; D018398:Homeodomain Proteins