Evaluation of the Differentiation Potential of Human Theca Stem Cells to Oocyte-Like Cells in the Ovary of Women of Different Reproductive Ages

Subject Areas : Journal of Animal Biology

Seyedeh Nasim Mirbahari ¹ , َAzam Dalman ² , Fatemeh Hasani ³ , Mehdi Totonchi ⁴

1 - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
2 - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
3 - Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
4 - Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

Received: 2022-04-12 Accepted : 2022-07-19 Published : 2023-02-20

Keywords: Ovary, Theca stem cells, In-vitro differentiation, Oocyte-like cells,

Abstract :

The evidence about the existence and function of ovarian stem cells (OSCs) is increasing. In a previous study, we isolated human theca stem cells (hTSCs) from the ovary of a 19-year-old patient and differentiated them into oocyte-like cells (hOLCs) under induced conditions. In this study, in order to prove the repeatability of this experiment as well as the presence of these cells in the ovaries of patients with higher reproductive age and their differentiation potential in-vitro, hTSCs were isolated from the ovaries of 20 and 38-year-old patients and their differentiation potential into hOLCs has been evaluated. In this experimental interventional study, based on the instructions of the previous study, hTSCs were isolated from small antral follicles with a size of 3 to 5 mm. These cells were cultured in six-well plates with A number of 5×104 cells per well in DMEM/F12 induction medium containing FBS, human follicular fluid, glutamine and pyruvate for 40 days. Then their development process was measured by assessing the morphology, size and viability. hTSCs were successfully isolated from ovarian tissue of 20 and 38-year-old patients and cultured in DMEM/F-12 medium containing growth factors (EGF, FGF, GDNF, etc.). After 12 days, hTSCs in both patients started to differentiate into hOLCs and their morphology changed from spindle-shaped to round. The size of hOLCs increased during the differentiation period in both patients (from 20-25 µm to 50 µm). The survival of hOLCs compared to hTSCs was similar in all three patients and did not differ significantly. hTSCs can be isolated from ovaries of women of different reproductive ages and there is no difference in their differentiation pattern to hOLCs in laboratory (in-vitro) conditions.

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