Generation of XXXX Embryonic Germ Cells via Inhibition of TGF   and MEK Pathways.

@entity9096 @entity7418 @entity1810 @entity35 @entity2008 @entity44
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 @entity35 EG cells with inhibition of @entity1810 ( @entity1810 ) and mitogen-activated protein kinase kinase (MEK) signaling pathways. MATERIALS AND METHODS: In this experimental study, @entity35 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 @entity7418 ( @entity7418 ) 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 @entity44 formation as well. Moreover, chromosomal stability of two different EG cells were assessed through G-banding technique. RESULTS: Formerly, derivation of @entity35 EG cells were observed solely in the presence of glycogen synthase kinase-3 ( @entity9096 ) and MEK pathway inhibitors. Due to some drawbacks of inhibiting @entity9096 molecules such as @entity2008 , in the present study we have attempted to assess a feeder-free protocol that derives EG cells by the simultaneous suppression of @entity1810 signaling and the MEK pathway. We have shown that @entity35 EG cells could be generated in the presence of two inhibitors that suppressed the above mentioned pathways. Of note, inhibition of @entity1810 instead of @entity9096 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. CONCLUSION: @entity35 EG cells with higher karyotype stability establish from PGCs by inhibiting @entity1810 and MEK signaling pathways.
