title: Influence of activin A supplementation during human embryonic stem cell derivation on germ cell differentiation potential.
abstract: Human embryonic stem cells (hESCs) are more similar to "primed" mouse epiblast stem cells (mEpiSCs). mEpiSCs, which are derived in Activin A, show an increased propensity to form primordial germ cell (PGC)-like cells in response to bone morphogenic protein 4 (BMP4). Hence, we hypothesized that hESCs derived in the presence of Activin A may be more competent in differentiating towards PGC-like cells after supplementation with BMP4 compared to standard hESC lines. We were able to successfully derive two hESC lines in the presence of Activin A, which were pluripotent and showed higher base levels of STELLA and cKIT compared to standard hESC lines derived without Activin A addition. Furthermore, upon differentiation as embryoid bodies in the presence of BMP4, we observed upregulation of VASA at day 7, both at the transcript and protein level compared to standard hESC lines, which appeared to take longer time for PGC specification. Unlike other hESC lines, nuclear pSMAD2/3 presence confirmed that Activin signalling was switched on in Activin A-derived hESC lines. They were also responsive to BMP4 based on nuclear detection of pSMAD1/5/8 and showed endodermal differentiation as a result of GATA-6 expression. Hence, our results provide novel insights into the impact of hESC derivation in the presence of Activin A and its subsequent influence on germ cell differentiation potential in vitro.
authors.affiliation: 1 Department for Reproductive Medicine, Ghent University Hospital , Ghent, Belgium .
authors.forename: Galbha
authors.initials: G
authors.lastname: Duggal
authors.forename: Björn
authors.initials: B
authors.lastname: Heindryckx
authors.forename: Sharat
authors.initials: S
authors.lastname: Warrier
authors.forename: Thomas
authors.initials: T
authors.lastname: O'Leary
authors.forename: Margot
authors.initials: M
authors.lastname: Van der Jeught
authors.forename: Sylvie
authors.initials: S
authors.lastname: Lierman
authors.forename: Liesbeth
authors.initials: L
authors.lastname: Vossaert
authors.forename: Tom
authors.initials: T
authors.lastname: Deroo
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.forename: Susana M
authors.initials: SM
authors.lastname: Chuva de Sousa Lopes
authors.forename: Petra
authors.initials: P
authors.lastname: De Sutter
mesh_terms: D028341:Activins / Q000494:pharmacology*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D058732:Embryoid Bodies / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004707:Endoderm / Q000187:drug effects / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D050993:GATA6 Transcription Factor / Q000378:metabolism; D020869:Gene Expression Profiling; D005854:Germ Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4; D050993:GATA6 Transcription Factor; D016212:Transforming Growth Factor beta; C422405:activin A; D028341:Activins