title: Generation of Rat Embryonic Germ Cells via Inhibition of TGFß and MEK Pathways.
abstract: OBJECTIVE
  Embryonic germ (EG) cells are the results of reprogramming primordial germ cells (PGC) in vitro. Studying these cells can be of benefit in determining the mechanism by which specialized cells acquire pluripotency. Therefore in the current study we have tried to derive rat EG cells with inhibition of transforming growth factor-β (TGFβ) and mitogen-activated protein kinase kinase (MEK) signaling pathways.
  
  
  METHODS
  In this experimental study, rat PGCs were cultured under feeder free condition with two small molecules that inhibited the above mentioned pathways. Under this condition only two-day presence of stem cell factor (SCF) as a survival factor was applied for PGC reprogramming. Pluripotency of the resultant EG cells were further confirmed by immunofluorescent staining, directed differentiation ability to neural and cardiac cells, and their contribution to teratoma formation as well. Moreover, chromosomal stability of two different EG cells were assessed through G-banding technique.
  
  
  RESULTS
  Formerly, derivation of rat EG cells were observed solely in the presence of glycogen synthase kinase-3 (GSK3β) and MEK pathway inhibitors. Due to some drawbacks of inhibiting GSK3β molecules such as increases in chromosomal aberrations, in the present study we have attempted to assess a feeder-free protocol that derives EG cells by the simultaneous suppression of TGFβ signaling and the MEK pathway. We have shown that rat EG cells could be generated in the presence of two inhibitors that suppressed the above mentioned pathways. Of note, inhibition of TGFβ instead of GSK3β significantly maintained chromosomal integrity. The resultant EG cells demonstrated the hallmarks of pluripotency in protein expression level and also showed in vivo and in vitro differentiation capacities.
  
  
  CONCLUSIONS
  Rat EG cells with higher karyotype stability establish from PGCs by inhibiting TGFβ and MEK signaling pathways.
authors.affiliation: Department of Physiology and Pharmacology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Mohammadi
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Farnoosh
authors.initials: F
authors.lastname: Attari
authors.affiliation: Department of Physiology and Pharmacology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
authors.forename: Vahab
authors.initials: V
authors.lastname: Babapour
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Seyedeh-Na Seh
authors.initials: SN
authors.lastname: Hassani
authors.affiliation: Department of Genetics at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Najmehsadat
authors.initials: N
authors.lastname: Masoudi
authors.affiliation: Department of Embryology at Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
authors.forename: Abdolhossein
authors.initials: A
authors.lastname: Shahverdi
authors.affiliation: Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran ; Department of Developmental Biology, University of Science and Culture, ACECR, Tehran, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Baharvand
keywords: Pluripotency; Rat; TGFβ Pathway