In VitroDifferentiation 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.

Analysis of Oocyte-Like Cells Differentiated from Porcine Fetal Skin-Derived Stem Cells

“…When placed in an appropriate female environment, PGCs were previously demonstrated to spontaneously proceed into the oocyte path in vivo [31,52]. In addition, male ES cells were shown to differentiate into oocytes in vitro [53]. In our experiment, the OLCs differentiated from male stem cells reached diameters and were morphologically very similar to OLCs derived from female stem cells.…”

Sertoli cell-conditioned medium induces germ cell differentiation in human embryonic stem cells

“…Induced germ cell differentiation protocols can be divided into two main groups. In one group of protocols, specific factors are added to the differentiating cells in order to increase germ cell derivation (BMP4 [5,[10][11][12], retinoic acid (RA) [7,[13][14][15], basic fibroblast growth factor (bFGF) [16], testicular cell conditioned medium [14,17], forskolin [15] and wingless-related MMTV integration site 3A (Wnt3a) [11]). Other protocols focus on the selection and enrichment of the spontaneously derived germ cell population [6,18,19].…”

Use of pluripotent stem cells for reproductive medicine: are we there yet?

“…However, these animals were only able to survive for five months, most likely due to impaired sex-specific imprinting in mESCderived male gametes (Nayernia et al 2006). Several other groups have tried different differentiation strategies and growth factor cocktails, but the imprinting status, germ cell maturation and meiosis in vitro remain difficult obstacles to produce functional mESC-derived gametes (Lacham-Kaplan et al 2006;Kerkis et al 2007;Qing et al 2007;Salvador et al 2008). The recent breakthrough work of Hayashi and colleagues has provided a promising future to the germ cell differentiation potential of ESCs (Hayashi et al 2011(Hayashi et al , 2012.…”