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In vitro and in vivo germ line potential of stem cells derived from newborn mouse skin.

Dyce PW, Liu J, Tayade C, Kidder GM, Betts DH, Li J - PLoS ONE (2011)

Bottom Line: Flow cytometric analysis revealed that ∼ 0.3% of the freshly isolated skin cells were GFP(+).The GFP-positive cells increased to ∼ 7% after differentiation, suggesting that the GFP(+) cells could be of in vivo origin, but are more likely induced upon being cultured in vitro.GFP(+) oocytes were identified within a subpopulation of follicles in the resulting growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada.

ABSTRACT
We previously reported that fetal porcine skin-derived stem cells were capable of differentiation into oocyte-like cells (OLCs). Here we report that newborn mice skin-derived stem cells are also capable of differentiating into early OLCs. Using stem cells from mice that are transgenic for Oct4 germline distal enhancer-GFP, germ cells resulting from their differentiation are expected to be GFP(+). After differentiation, some GFP(+) OLCs reached 40-45 µM and expressed oocyte markers. Flow cytometric analysis revealed that ∼ 0.3% of the freshly isolated skin cells were GFP(+). The GFP-positive cells increased to ∼ 7% after differentiation, suggesting that the GFP(+) cells could be of in vivo origin, but are more likely induced upon being cultured in vitro. To study the in vivo germ cell potential of skin-derived cells, they were aggregated with newborn ovarian cells, and transplanted under the kidney capsule of ovariectomized mice. GFP(+) oocytes were identified within a subpopulation of follicles in the resulting growth. Our finding that early oocytes can be differentiated from mice skin-derived cells in defined medium may offer a new in vitro model to study germ cell formation and oogenesis.

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Formation of oocytes in vivo following transplantation of cell aggregates into immunodeficient mice.A. Growth under the kidney capsule (arrow) following 16 weeks of grafting. B. A follicle containing a GFP-positive oocyte (black arrow) and two follicles containing GFP-negative oocytes (white arrows). C. A follicle with a GFP-negative oocyte. D, E. Follicles containing positive oocytes (black arrows). Scale bars: B, 120 µm, C–E, 60 µm.
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pone-0020339-g008: Formation of oocytes in vivo following transplantation of cell aggregates into immunodeficient mice.A. Growth under the kidney capsule (arrow) following 16 weeks of grafting. B. A follicle containing a GFP-positive oocyte (black arrow) and two follicles containing GFP-negative oocytes (white arrows). C. A follicle with a GFP-negative oocyte. D, E. Follicles containing positive oocytes (black arrows). Scale bars: B, 120 µm, C–E, 60 µm.

Mentions: To study the in vivo germ cell potential of the newborn mouse skin-derived stem cells, we mixed 0.3×106 stem cells with 0.6×106 newborn ovarian cells and cultured 72 hours in the differentiation medium. Following culture, the reaggregated cells were transplanted under the kidney capsules of immunodeficient Rag2−/−,gamma(c)−/− ovariectomized mice. The kidney was chosen as the host organ of the transplant as it is one of the most heavily vascularized organs in the body and vascularization of the graft occurs within a short period of time so that the graft responds to the hormonal milieu of the host. Growths were identified under the transplanted kidney capsule after in vivo incubation (Figure 8A). Follicles with GFP-positive or negative oocytes, detected by immunostaining, were found within the growth (Figure 8B). Figure 8C is a follicle with GFP-negative oocyte, while Figures 8D and E show follicles with GFP-positive oocytes. The presence of a GFP-positive oocyte in the reconstituted transplanted ovarian structures provides evidence that under in vivo conditions, the newborn mouse skin-derived stem cells have the potential to form oocytes.


In vitro and in vivo germ line potential of stem cells derived from newborn mouse skin.

Dyce PW, Liu J, Tayade C, Kidder GM, Betts DH, Li J - PLoS ONE (2011)

Formation of oocytes in vivo following transplantation of cell aggregates into immunodeficient mice.A. Growth under the kidney capsule (arrow) following 16 weeks of grafting. B. A follicle containing a GFP-positive oocyte (black arrow) and two follicles containing GFP-negative oocytes (white arrows). C. A follicle with a GFP-negative oocyte. D, E. Follicles containing positive oocytes (black arrows). Scale bars: B, 120 µm, C–E, 60 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3101249&req=5

pone-0020339-g008: Formation of oocytes in vivo following transplantation of cell aggregates into immunodeficient mice.A. Growth under the kidney capsule (arrow) following 16 weeks of grafting. B. A follicle containing a GFP-positive oocyte (black arrow) and two follicles containing GFP-negative oocytes (white arrows). C. A follicle with a GFP-negative oocyte. D, E. Follicles containing positive oocytes (black arrows). Scale bars: B, 120 µm, C–E, 60 µm.
Mentions: To study the in vivo germ cell potential of the newborn mouse skin-derived stem cells, we mixed 0.3×106 stem cells with 0.6×106 newborn ovarian cells and cultured 72 hours in the differentiation medium. Following culture, the reaggregated cells were transplanted under the kidney capsules of immunodeficient Rag2−/−,gamma(c)−/− ovariectomized mice. The kidney was chosen as the host organ of the transplant as it is one of the most heavily vascularized organs in the body and vascularization of the graft occurs within a short period of time so that the graft responds to the hormonal milieu of the host. Growths were identified under the transplanted kidney capsule after in vivo incubation (Figure 8A). Follicles with GFP-positive or negative oocytes, detected by immunostaining, were found within the growth (Figure 8B). Figure 8C is a follicle with GFP-negative oocyte, while Figures 8D and E show follicles with GFP-positive oocytes. The presence of a GFP-positive oocyte in the reconstituted transplanted ovarian structures provides evidence that under in vivo conditions, the newborn mouse skin-derived stem cells have the potential to form oocytes.

Bottom Line: Flow cytometric analysis revealed that ∼ 0.3% of the freshly isolated skin cells were GFP(+).The GFP-positive cells increased to ∼ 7% after differentiation, suggesting that the GFP(+) cells could be of in vivo origin, but are more likely induced upon being cultured in vitro.GFP(+) oocytes were identified within a subpopulation of follicles in the resulting growth.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada.

ABSTRACT
We previously reported that fetal porcine skin-derived stem cells were capable of differentiation into oocyte-like cells (OLCs). Here we report that newborn mice skin-derived stem cells are also capable of differentiating into early OLCs. Using stem cells from mice that are transgenic for Oct4 germline distal enhancer-GFP, germ cells resulting from their differentiation are expected to be GFP(+). After differentiation, some GFP(+) OLCs reached 40-45 µM and expressed oocyte markers. Flow cytometric analysis revealed that ∼ 0.3% of the freshly isolated skin cells were GFP(+). The GFP-positive cells increased to ∼ 7% after differentiation, suggesting that the GFP(+) cells could be of in vivo origin, but are more likely induced upon being cultured in vitro. To study the in vivo germ cell potential of skin-derived cells, they were aggregated with newborn ovarian cells, and transplanted under the kidney capsule of ovariectomized mice. GFP(+) oocytes were identified within a subpopulation of follicles in the resulting growth. Our finding that early oocytes can be differentiated from mice skin-derived cells in defined medium may offer a new in vitro model to study germ cell formation and oogenesis.

Show MeSH
Related in: MedlinePlus