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Isolation, culture, and characterization of primordial germ cells in Mongolian sheep

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Abstract

This study was performed to culture and preliminarily identify the primordial germ cells (PGCs) isolated from the genital ridge of the Mongolian sheep fetus. The growth characteristics of the sheep PGCs were detected in different culture systems such as culture media, resources, and state and passages of feeder cells. The obtained embryonic germ (EG) cells were identified by morphology, enzymology, and immunofluorescence. The results showed that the sheep EG cell colonies were ridgy, typically nest like, and compact, and had regular edges. Alkaline phosphatase staining reaction was weakly positive. EG cells expressed Kit, Rex-1, Nanog, and Oct-4. Immunofluorescence detection was weakly positive for Oct3/4, whereas positive for SSEA-1, SSEA-3, SSEA-4, TRA-1-61, and TRA-1-80.

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References

  • Bao L.; He L.; Chen J.; Wu Z.; Liao J.; Rao L.; Ren J.; Li H.; Zhu H.; Qian L.; Gu Y.; Dai H.; Xu X.; Zhou J.; Wang W.; Cui C.; Xiao L. Reprogramming of ovine adult fibroblasts to pluripotency via drug-inducible expression of defined factors. Cell Res. 21: 600–608; 2011.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Dattena M.; Chessa B.; Lacerenza D.; Accardo C.; Pilichi S.; Mara L.; Chessa F.; Vincenti L.; Cappai P. Isolation, culture, and characterization of embryonic cell lines from vitrified sheep blastocysts. Molecular Reproduction and Development 73: 31–39; 2006.

    Article  PubMed  CAS  Google Scholar 

  • De Felici M.; McLaren A. Isolation of mouse primordial germ cells. Exp Cell Res 142: 476–482; 1982.

    Article  PubMed  Google Scholar 

  • De Miguel M. P.; Kerr C. L.; López-Iglesias P.; Alcaina Y. Techniques and conditions for embryonic germ cell derivation and culture. In: Kallos M. (ed) Embryonic stem cells—basic biology to bioengineering. InTech, Vienna, pp 425–448; 2011.

    Google Scholar 

  • Hall V. Porcine embryonic stem cells: a possible source for cell replacement therapy. Stem Cell Reviews and reports 4: 275–282; 2008.

    Article  PubMed  Google Scholar 

  • Insausti C. L.; Blanquer M.; Bleda P.; Iniesta P.; Majado M. J.; Castellanos G.; Moraleda J. M. The amniotic membrane as a source of stem cells. Histol Histopathol 25: 91–98; 2010.

    PubMed  CAS  Google Scholar 

  • Jiang F. X. Purification and characterization of primordial germ cells in male rat fetuses. Ital J Anat Embryol 103: 31–39; 1998.

    PubMed  CAS  Google Scholar 

  • Kakegawa R.; Teramura T.; Takehara T.; Anzai M.; Mitani T.; Matsumoto K.; Saeki K.; Sagawa N.; Fukuda K.; Hosoi Y. Isolation and culture of rabbit primordial germ cells. J Reprod Dev 54: 352–357; 2008.

    Article  PubMed  CAS  Google Scholar 

  • Kühholzer B.; Baguisi A.; Overstrom E. W. Long-term culture and characterization of goat primordial germ cells. Theriogenology 53: 1071–1079; 2000.

    Article  PubMed  Google Scholar 

  • Ledda S.; Bogliolo L.; Bebbere D.; Ariu F.; Pirino S. Characterization, isolation and culture of primordial germ cells in domestic animals: recent progress and insights from the ovine species. Theriogenology 74: 534–543; 2010.

    Google Scholar 

  • Ledda S.; Naitana S.; Loi P.; Cappai P.; Moor R. M.; Calvia P.; Fulka J. Transplantation of primordial germ cells (PGCs) into enucleated oocytes in sheep. Theriogenology 45: 288; 1996.

    Google Scholar 

  • Lee C. K.; Scales N.; Newton G.; Piedrahita J. A. Isolation and initial characterization of primordial germ cell (PGC)—derived cells from goat, rabbit and rats. Theriogenology 49: 388; 1998.

    Article  Google Scholar 

  • Li Y.; Cang M.; Lee A. S.; Zhang K.; Liu D. Reprogramming of sheep fibroblasts into pluripotency under a drug-inducible expression of mouse-derived defined factors. Plos One 6: e15947; 2011.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Matsui Y.; Zsebo K.; Hogan B. L. Derivation of pluripotential embryonic stem cells from murine primordial germ cells in cultures. Cell 70: 841–847; 1992.

    Article  PubMed  CAS  Google Scholar 

  • Miki T.; Lehmann T.; Cai H.; Stolz D. B.; Strom S. C. Stem cell characteristics of amniotic epithelial cells. Stem Cells 23: 1549–1559; 2005.

    Google Scholar 

  • Miki T.; Strom S. C. Amnion-derived pluripotent/pluripotent stem cells. Stem Cell Rev 2: 133–142; 2006.

    Google Scholar 

  • Moreno-Ortiz H.; Esteban-Perez C.; Badran W.; Kent-First M. Isolation and derivation of mouse embryonic germinal cells. Journal of Visualized Experiments 32: e1635; 2009.

    Google Scholar 

  • Parolini O.; Alviano F.; Bagnara G. P.; Bilic G.; Bühring H. J.; Evangelista M.; Hennerbichler S.; Liu B.; Magatti M.; Mao N.; Miki T.; Marongiu F.; Nakajima H.; Nikaido T.; Portmann-Lanz C. B.; Sankar V.; Soncini M.; Stadler G.; Surbek D.; Takahashi T. A.; Redl H.; Sakuragawa N.; Wolbank S.; Zeisberger S.; Zisch A.; Strom S. C. Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells. Stem cell 26: 300–311; 2008.

    Article  Google Scholar 

  • Shamblott M. J.; Axelman J.; Wang S.; Bugg E. M.; Littlefield J. W.; Donovan P. J.; Blumenthal P. D.; Huggins G. R.; Gearhart J. D. Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc. Natl Acad Sci 95: 13726–13731; 1998.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  • Shim H.; Gutiérrez-Adán A.; Chen L. R.; BonDurant R. H.; Behboodi E.; Anderson G. B. Isolation of pluripotent stem cells from cultured porcine primordial germ cells. Biology of Reproduction 57: 1089–1095; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Wrobel K. H.; Süß F. Identification and temporospatial distribution of bovine primordial germ cells prior to gonadal sexual differentiation. Anat Embryol 197: 451–467; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Yu J.; Thomson J. A. Pluripotent stem cell lines. Genes Dev 22: 1987–1997; 2008.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

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Acknowledgments

Funding was supplied by the Chinese National 863 Project (2008AA101005). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Correspondence to Huan Min Zhou.

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Editor: T. Okamoto

Chun Xia Liu, Wen Long Wang, and Rui Yuan Zhao contributed to the research equally.

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Liu, C.X., Wang, W.L., Zhao, R.Y. et al. Isolation, culture, and characterization of primordial germ cells in Mongolian sheep. In Vitro Cell.Dev.Biol.-Animal 50, 207–213 (2014). https://doi.org/10.1007/s11626-013-9695-0

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  • DOI: https://doi.org/10.1007/s11626-013-9695-0

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