title: In vitro differentiation of male mouse embryonic stem cells into both presumptive sperm cells and oocytes.
abstract: Pioneer work in male mouse embryonic stem (ES) cells differentiation into germ cells (GC) showed generations of male or female gametes in separate experiments, using genetically manipulated or preselected ES cells. In an attempt to produce both types of gametes from male mouse ES cells without any genetic manipulation or preselection, we induce the differentiation by retinoic acid (RA) within nonadherent embryoid bodies (EB). It seems that gamete-like cell formation occurs in the correct manner based on the expression of early and late GC-specific genes such as Oct-4, Mvh, Stella, Dazl, Piwil 2, Pdrd 1, Rex 14, Rnf 17, Bmp8b, Acrosin, Stra-8, Haprin, LH-R, Gdf9, Zp3, Zp2, Sycp1, and Sycp3. Immunofluorescence analysis of morphologically well-formed GC and presumptive gametes showed positive labeling for SSEA1, Oct-4, EMA-1, FE-J1, Dazl, Fragilis, Mvh, Acrosin, and acetylated alpha-tubulin. Conventional cytogenetic and FISH analysis indicated a chromosome reduction in ES-derived GC. Our data suggest that ES cells with XY chromosomes can produce under the same experimental conditions both types of presumptive gametes, and this production depends on their positional and temporal information within the EB context.
authors.affiliation: Clínica e Centro de Pesquisa em Reprodução Humana Roger Abdelmassih, São Paulo, SP, Brasil. akerkis@usp.br
authors.forename: Alexandre
authors.initials: A
authors.lastname: Kerkis
authors.forename: Simone A S
authors.initials: SA
authors.lastname: Fonseca
authors.forename: Rui C
authors.initials: RC
authors.lastname: Serafim
authors.forename: Thais M C
authors.initials: TM
authors.lastname: Lavagnolli
authors.forename: Soraya
authors.initials: S
authors.lastname: Abdelmassih
authors.forename: Roger
authors.initials: R
authors.lastname: Abdelmassih
authors.forename: Irina
authors.initials: I
authors.lastname: Kerkis
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D019976:Cloning, Organism / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology*; D005821:Genetic Techniques; D006728:Hormones / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D013094:Spermatozoa / Q000166:cytology*; D014212:Tretinoin / Q000378:metabolism
chemical_list: D006728:Hormones; D014212:Tretinoin