Zdravniški Vestnik (Feb 2018)

OVARIAN INFERTILITY, STEM CELLS AND OOGENSIS IN VITRO

  • Irma Virant-Klun,
  • Andrej Vogler,
  • Jasna Šinkovec,
  • Matjaž Hren,
  • Kristina Gruden,
  • Elvira Maličev,
  • Helena Meden-Vrtovec

DOI
https://doi.org/10.6016/ZdravVestn.2737
Journal volume & issue
Vol. 78, no. 0

Abstract

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Background: In different human adult tissues and organs embryonic-like stem cells have already been found. A question is whether they are present also in the adult human ovary. In this study we tried to find whether embryonic-like stem cells are present in the ovarian surface epithe- lium of women with premature ovarian failure, and whether these cells can differentiate into oocytes by in vitro culture. Methods: In 10 women with premature ovarian failure the ovarian surface epithelium was laparo- scopically scraped. We tried to immunomagnetically isolate embryonic-like stem cells from the suspension of scraped cells and to confirm their existence immunocytochemically by the expression of the markers c-kit and surface antigen SSEA-4, characteristic of pluripotent stem cells. In the lab, we tried to differentiate stem cells into oocytes in a cell culture medium with added heterologous follicular fluid from the in vitro fertilization programme, which is rich in substances, and important for oocyte growth and maturation. The differentiation of oocyte-like cells was screened. Oocyte-like cells were isolated from the cell culture and im- munocytochemically stained for oocyte-specific markers (c-kit, VASA, ZP2-zona pellucida). Oocyte-like cells were analyzed for the expression of oocyte-specific genes (Oct-4A, ZP3, c-kit, SCP-3) by single-cell PCR. Results: From the ovarian surface epithelium of all patients small yellow coloured cells with diam- eters of 2 to 4 μm and positive for markers of pluripotent stem cells were isolated. In the lab, these cells were cultured into oocyte-like cells with diameter ranging from 60 to 80 μm. Some of oocyte-like cells were positive for oocyte-specific markers and expressed the genes, characteristic of oocytes, including meiotic genes. Conclusions: In patients with premature ovarian failure it is possible to culture oocyte-like cells from ovarian surface epithelium stem cells. Intense research is needed to evaluate the quality of oocyte-like cells developed in vitro and to estimate their potential use for in vitro fertilization in the far future.

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