Limits...
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.

Show MeSH

Related in: MedlinePlus

Detection of meiosis markers in OLCs by RT-PCR.A. Expression of Dmc1, Scp3 and lack of expression of Rec8 in the OLCs. Rpii was used as a loading control. B. Germinal vesicle-like structure (left, arrow) in an OLC similar to that of a natural oocyte (right).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3101249&req=5

pone-0020339-g007: Detection of meiosis markers in OLCs by RT-PCR.A. Expression of Dmc1, Scp3 and lack of expression of Rec8 in the OLCs. Rpii was used as a loading control. B. Germinal vesicle-like structure (left, arrow) in an OLC similar to that of a natural oocyte (right).

Mentions: Next we sought to study if these OLCs can undergo meiosis, a function which is unique to oocytes and male germ cells at later stages of their development. During meiosis REC8 is involved in chromosome reduction and segregation and co-localizes with SPC3 and DMC1 (Tarsounas, Morita et al. 1999). RT-PCR revealed that the transcript of Rec8 was expressed in the Ov group, as well as the oocytes from the ovary of newborn mice but not in OLCs. Scp3 and Dmc1 transcripts are present in the OLCs and the two positive controls (Figure 7A). Immunocytochemistry of SCP3 was performed on chromosome spreads. It was found that the vast majority of SCP3-stained cells appeared to show discontinuous staining patterns that do not resemble any typical prophase stage (data not shown). Morphologically, some OLCs appeared to have a germinal vesicle (GV) like structure (Figure 7B). These data suggest some of the OLCs derived from SC can initiate meiosis, but they failed to progress beyond the MI stage in our current culture environment.


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)

Detection of meiosis markers in OLCs by RT-PCR.A. Expression of Dmc1, Scp3 and lack of expression of Rec8 in the OLCs. Rpii was used as a loading control. B. Germinal vesicle-like structure (left, arrow) in an OLC similar to that of a natural oocyte (right).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0020339-g007: Detection of meiosis markers in OLCs by RT-PCR.A. Expression of Dmc1, Scp3 and lack of expression of Rec8 in the OLCs. Rpii was used as a loading control. B. Germinal vesicle-like structure (left, arrow) in an OLC similar to that of a natural oocyte (right).
Mentions: Next we sought to study if these OLCs can undergo meiosis, a function which is unique to oocytes and male germ cells at later stages of their development. During meiosis REC8 is involved in chromosome reduction and segregation and co-localizes with SPC3 and DMC1 (Tarsounas, Morita et al. 1999). RT-PCR revealed that the transcript of Rec8 was expressed in the Ov group, as well as the oocytes from the ovary of newborn mice but not in OLCs. Scp3 and Dmc1 transcripts are present in the OLCs and the two positive controls (Figure 7A). Immunocytochemistry of SCP3 was performed on chromosome spreads. It was found that the vast majority of SCP3-stained cells appeared to show discontinuous staining patterns that do not resemble any typical prophase stage (data not shown). Morphologically, some OLCs appeared to have a germinal vesicle (GV) like structure (Figure 7B). These data suggest some of the OLCs derived from SC can initiate meiosis, but they failed to progress beyond the MI stage in our current culture environment.

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