Abstract
Infertility has long been a difficult issue for many couples. The successful differentiation of germ cells and live progeny from pluripotent stem cells brings new hope to the couples suffering with infertility. Here we successfully isolated human fetus skin-derived stem cells (hfSDSCs) from fetus skin tissue and demonstrated that hfSDSCs can be differentiated into early human germ cell-like cells (hGCLCs). These cells express human germ cell markers DAZL and VASA. Moreover, these pluripotent stem cell-derived hGCLCs are free of exogenous gene integration. When hfSDSCs were differentiated in porcine follicle fluid (PFF) conditioned media, which has been shown to promote the differentiation of mouse and porcine SDSCs into oocyte-like cells (OLCs), we observed some vesicular structures formed from hfSDSCs. Moreover, when hfSDSCs were cultured with specific conditioned media, we observed punctate and elongated SCP3 staining foci, indicating the initiation of meiosis. Ploidy analysis and fluorescent in situ hybridization (FISH) analysis indicated that a small percentage of putative 1N populations formed from hfSDSCs when compared with positive controls. In conclusion, our data here, for the first time, demonstrated that hfSDSCs possess the differentiation potential into germ lines, and they may differentiate both male and female hGCLCs in vitro under appropriate conditions.
Highlights
Public concerns on the application of induced pluripotent stem (iPS) cells mainly focus on the tumorigenicity and immunogenicity of transplanted iPS cells[8,9]
The skin tissues were trypsinized with TypLE Express for 15–30 min at 37 °C according to the gestational age and hfSDSCs were harvested as previously described[10]
embryoid body (EB)-like colonies formed from hfSDSCs exhibited similar morphology as those formed from human ES cell lines
Summary
Public concerns on the application of iPS cells mainly focus on the tumorigenicity and immunogenicity of transplanted iPS cells[8,9]. Skin-derived stem cells (SDSCs) from porcine or mouse show the ability to give rise to germ cell-like cells (GCLCs) even without reprogramming into the iPS cell stage[10,11,12]. These SDSC-derived germ cells express germ cell markers and show similar DNA methylation patterns to that of their in vivo counterparts[12]. We demonstrated that hfSDSCs showed the potential for germline differentiation in vitro, which may provide a new model for elucidating mechanisms underlying human gametogenesis
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