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Article
Analysis of Oocyte-Like Cells Differentiated from Porcine Fetal Skin-Derived Stem Cells
Stem Cells and Development (2011)
  • Paul W. Dyce, University of Guelph
  • Wei Shen, University of Guelph
  • Evanna Huynh, University of Guelph
  • Hua Shao, University of Guelph
  • Daniel A. F. Villagómez
  • Gerald M. Kidder, The University of Western Ontario
  • W Allan King, University of Guelph
  • Julang Li, University of Guelph
Abstract

We previously reported the differentiation of cells derived from porcine female fetal skin into cells resembling germ cells and oocytes. A subpopulation of these cells expressed germ cell markers and formed aggregates resembling cumulus-oocyte complexes. Some of these aggregates extruded large oocyte-like cells (OLCs) that expressed markers consistent with those of oocytes. The objective of the current study was to further characterize OLCs differentiated from porcine skin-derived stem cells. Reverse transcriptase (RT)-polymerase chain reaction and Western blot revealed the expression of connexin37 and connexin43, both of which are characteristic of ovarian follicles. The expression of meiosis markers DMC1 and synaptonemal complex protein, but not STRA8 and REC8, was detected in the OLC cultures. Immunofluorescence with an antibody against synaptonemal complex protein on chromosome spreads revealed a very small subpopulation of stained OLCs that had a similar pattern to leptotene, zytotene, or pachytene nuclei during prophase I of meiosis. Sodium bisulfite sequencing of the differentially methylated region of H19 indicated that this region is almost completely demethylated in OLCs, similar to in vivo-derived oocytes. We also investigated the differentiation potential of male skin-derived stem cells in the same differentiation medium. Large cells with oocyte morphology were generated in the male stem cell differentiation cultures. These OLCs expressed oocyte genes such as octamer-binding transcription factor 4 (OCT4), growth differentiation factor-9b (GDF9B), deleted in azoospermia-like (DAZL), VASA, zona pellucida B (ZPB), and zona pellucida C (ZPC). It was concluded that skin-derived stem cells from both male and female porcine fetuses are capable of entering an oocyte differentiation pathway, but the culture system currently in place is inadequate to support the complete development of competent oocytes.

Keywords
  • Stem cell,
  • Reproduction,
  • Oocyte-like cell
Publication Date
May, 2011
Publisher Statement
This is a copy of an article published in Stem Cells and Development © 2011 Mary Ann Liebert, Inc.; Stem Cells and Development is available online at: http://www.liebertonline.com.
Citation Information
Paul W. Dyce, Wei Shen, Evanna Huynh, Hua Shao, et al.. "Analysis of Oocyte-Like Cells Differentiated from Porcine Fetal Skin-Derived Stem Cells" Stem Cells and Development Vol. 20 Iss. 5 (2011)
Available at: http://works.bepress.com/pauldyce/5/