Limits...
Differentiation of early germ cells from human skin-derived stem cells without exogenous gene integration.

Ge W, Ma HG, Cheng SF, Sun YC, Sun LL, Sun XF, Li L, Dyce P, Li J, Shi QH, Shen W - Sci Rep (2015)

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Reproductive Sciences, Qingdao Agricultural University, Qingdao, Shandong 266109, China.

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.

No MeSH data available.


Related in: MedlinePlus

Differentiation of hfSDSCs into human germ cell-like cells (hGCLCs) in porcine follicle fluid (PFF) conditioned media.(A) DAZL- and VASA-positive hGCLCs. (B) Differentiation of human, porcine and mouse SDSCs in PFF conditioned media. (C) Expression of germ cell and meiosis markers including DAZL, VASA, SCP3 and AMH in hfSDSCs-differentiated cells using PFF conditioned media.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4561906&req=5

f2: Differentiation of hfSDSCs into human germ cell-like cells (hGCLCs) in porcine follicle fluid (PFF) conditioned media.(A) DAZL- and VASA-positive hGCLCs. (B) Differentiation of human, porcine and mouse SDSCs in PFF conditioned media. (C) Expression of germ cell and meiosis markers including DAZL, VASA, SCP3 and AMH in hfSDSCs-differentiated cells using PFF conditioned media.

Mentions: Since our previous study demonstrated that fetal porcine and newborn mouse SDSCs have the intrinsic ability to differentiate into GCLCs101112, we next verified whether hfSDSCs can also be induced into hGCLCs in vitro by using the same protocol. Immunofluorescence data revealed that some subpopulations of the hfSDSCs exhibited positive expression of hGCLCs biomarkers including DAZL (Deleted in Azoospermia-like) and VASA (Fig. 2A). Previous study reported that shiny colonies of porcine or mouse GCLCs are morphologically distinctive after 15–30 days of differentiation101112, interestingly, the hGCLCs did not show morphological distinction from other cells (Fig. 2B). hGCLCs aggregated with peripheral cells and cannot be distinguished from them via morphology. Although oocyte-like cells (OLCs) differentiated from SDSCs were observed in the mouse and porcine models, we did not observe typical OLCs using the same method. However, the vesicular structures were observed when hfSDSCs were differentiated with PFF conditioned media (indicated by red arrow) (Fig. 2B). Surprisingly, we unexpectedly observed that granulosa cell marker AMH (Anti-Mullerian Hormone) was upregulated in the vesicular structures, together with the elevated expression of germ cells markers and meiosis markers including DAZL, VASA and SCP3 (Synaptonemal Complex Protein 3) (Fig. 2C and Supplementary Fig. S2A). These indicate that these structures may be similar to the OLCs seen in the mouse and porcine models.


Differentiation of early germ cells from human skin-derived stem cells without exogenous gene integration.

Ge W, Ma HG, Cheng SF, Sun YC, Sun LL, Sun XF, Li L, Dyce P, Li J, Shi QH, Shen W - Sci Rep (2015)

Differentiation of hfSDSCs into human germ cell-like cells (hGCLCs) in porcine follicle fluid (PFF) conditioned media.(A) DAZL- and VASA-positive hGCLCs. (B) Differentiation of human, porcine and mouse SDSCs in PFF conditioned media. (C) Expression of germ cell and meiosis markers including DAZL, VASA, SCP3 and AMH in hfSDSCs-differentiated cells using PFF conditioned media.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561906&req=5

f2: Differentiation of hfSDSCs into human germ cell-like cells (hGCLCs) in porcine follicle fluid (PFF) conditioned media.(A) DAZL- and VASA-positive hGCLCs. (B) Differentiation of human, porcine and mouse SDSCs in PFF conditioned media. (C) Expression of germ cell and meiosis markers including DAZL, VASA, SCP3 and AMH in hfSDSCs-differentiated cells using PFF conditioned media.
Mentions: Since our previous study demonstrated that fetal porcine and newborn mouse SDSCs have the intrinsic ability to differentiate into GCLCs101112, we next verified whether hfSDSCs can also be induced into hGCLCs in vitro by using the same protocol. Immunofluorescence data revealed that some subpopulations of the hfSDSCs exhibited positive expression of hGCLCs biomarkers including DAZL (Deleted in Azoospermia-like) and VASA (Fig. 2A). Previous study reported that shiny colonies of porcine or mouse GCLCs are morphologically distinctive after 15–30 days of differentiation101112, interestingly, the hGCLCs did not show morphological distinction from other cells (Fig. 2B). hGCLCs aggregated with peripheral cells and cannot be distinguished from them via morphology. Although oocyte-like cells (OLCs) differentiated from SDSCs were observed in the mouse and porcine models, we did not observe typical OLCs using the same method. However, the vesicular structures were observed when hfSDSCs were differentiated with PFF conditioned media (indicated by red arrow) (Fig. 2B). Surprisingly, we unexpectedly observed that granulosa cell marker AMH (Anti-Mullerian Hormone) was upregulated in the vesicular structures, together with the elevated expression of germ cells markers and meiosis markers including DAZL, VASA and SCP3 (Synaptonemal Complex Protein 3) (Fig. 2C and Supplementary Fig. S2A). These indicate that these structures may be similar to the OLCs seen in the mouse and porcine models.

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Reproductive Sciences, Qingdao Agricultural University, Qingdao, Shandong 266109, China.

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.

No MeSH data available.


Related in: MedlinePlus