title: Efficient induction of pluripotency in primordial germ cells by dual inhibition of TGF-β and ERK signaling pathways.
abstract: Primordial germ cells (PGCs) have the ability to be reprogrammed into a pluripotent state and are defined as embryonic germ cells (EGCs) in vitro. EGC formation is more efficient, has a shorter culture period than somatic cell reprogramming, and does not require exogenous genetic manipulation. Therefore, EGCs are a good model to analyze mechanisms by which committed cells acquire a pluripotent state. In the present study we have attempted to elucidate a more defined and robust protocol that promotes EGC generation through the suppression of transforming growth factor-β (TGF-β) and extracellular signal-regulated kinase (ERK) signaling pathways by SB431542 (SB) and PD0325901 (PD), respectively. Under this condition the efficiency of transformation of PGCs into EGCs was more than the inhibition of glycogen synthase kinase 3 and ERK signaling pathways. Pluripotency of the resultant-derived EGC lines were further confirmed by gene expression, immunofluorescent staining, directed differentiation ability, teratoma formation, and their contribution to chimeric mice and germ-line transmission. These results showed that PGCs from different embryonic stages (E8.5 and E12.5) could be reprogrammed, maintained, and expanded efficiently under feeder- and serum-free chemically defined conditions by dual inhibition of TGF-β and ERK signaling pathways, regardless of the animal's genetic background.
authors.affiliation: 1 Department of Animal Biology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran , Tehran, Iran .
authors.forename: Farnoosh
authors.initials: F
authors.lastname: Attari
authors.forename: Houri
authors.initials: H
authors.lastname: Sepehri
authors.forename: Hassan
authors.initials: H
authors.lastname: Ansari
authors.forename: Seyedeh-Nafiseh
authors.initials: SN
authors.lastname: Hassani
authors.forename: Fereshteh
authors.initials: F
authors.lastname: Esfandiari
authors.forename: Behrouz
authors.initials: B
authors.lastname: Asgari
authors.forename: Abdolhossein
authors.initials: A
authors.lastname: Shahverdi
authors.forename: Hossein
authors.initials: H
authors.lastname: Baharvand
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D004149:Dioxoles / Q000494:pharmacology; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology / Q000196:embryology; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D016212:Transforming Growth Factor beta; C506614:mirdametinib; D004159:Diphenylamine