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Adipose tissue–derived stem cells protect the primordial follicle pool from both direct follicle death and abnormal activation after ovarian tissue transplantation

  • Reproductive Physiology and Disease
  • Published:
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Abstract

Purpose

To investigate whether adipose tissue–derived stem cells (ASCs) protect the primordial follicle pool, not only by decreasing direct follicle loss but also by modulating follicle activation pathways.

Methods

Twenty nude mice were grafted with frozen-thawed human ovarian tissue from 5 patients. Ten mice underwent standard ovarian tissue transplantation (OT group), while the remaining ten were transplanted with ASCs and ovarian tissue (2-step/ASCs+OT group). Ovarian grafts were retrieved on days 3 (n = 5) and 10 (n = 5). Analyses included histology (follicle count and classification), immunohistochemistry (c-caspase-3 for apoptosis and LC3B for autophagy), and immunofluorescence (FOXO1 for PI3K/Akt activation and YAP for Hippo pathway disruption). Subcellular localization was determined in primordial follicles on high-resolution images using structured illumination microscopy.

Results

The ASCs+OT group showed significantly higher follicle density than the OT group (p = 0.01). Significantly increased follicle atresia (p < 0.001) and apoptosis (p = 0.001) were observed only in the OT group. In primordial follicles, there was a significant shift in FOXO1 to a cytoplasmic localization in the OT group on days 3 (p = 0.01) and 10 (p = 0.03), indicating follicle activation, while the ASCs+OT group and non-grafted controls maintained nuclear localization, indicating quiescence. Hippo pathway disruption was encountered in primordial follicles irrespective of transplantation, with nuclear YAP localized in their granulosa cells.

Conclusion

We demonstrate that ASCs exert positive effects on the ovarian reserve, not only by protecting primordial follicles from direct death but also by maintaining their quiescence through modulation of the PI3K/Akt pathway.

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Funding

This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (F.R.S.-FNRS/FRIA FC29657 awarded to Luciana Cacciottola, FNRS-PDR Convention T.0077.14, the Excellence of Science FNRS–EOS number 30443682, and grant 5/4/150/5 awarded to Marie-Madeleine Dolmans), Fonds Spéciaux de Recherche, Fondation St Luc and the Foundation Against Cancer, and donations from the Ferrero family.

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Authors and Affiliations

  1. Gynecology Research Unit, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. Mounier 52, bte B1.52.02, 1200, Brussels, Belgium

    Luciana Cacciottola, Thu Y. T. Nguyen, Camille Hossay & Marie-Madeleine Dolmans

  2. IREC Imaging Platform, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Av. Hippocrate 55, 1200, Brussels, Belgium

    Guillaume E. Courtoy

  3. Society for Research into Fertility, Av. Grandchamp 143, 1150, Brussels, Belgium

    Jacques Donnez

  4. Department of Gynecology, Cliniques Universitaires Saint-Luc, Av. Hippocrate 10, 1200, Brussels, Belgium

    Marie-Madeleine Dolmans

Authors
  1. Luciana Cacciottola
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  2. Guillaume E. Courtoy
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  3. Thu Y. T. Nguyen
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  4. Camille Hossay
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  5. Jacques Donnez
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  6. Marie-Madeleine Dolmans
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Correspondence to Marie-Madeleine Dolmans.

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Supplementary information

Supplementary Figure 1

– Positive and negative immunostaining controls. Positive controls were conducted using spleen tissue for caspase-3 (A), adrenal gland tissue for LC3B (B), bowel tissue for FOXO1 (C) and breast cancer tissue for YAP (D). Negative controls were obtained using non-specific antibodies. Scale bar: 50 μm. (PNG 44.0 mb).

High resolution image (TIF 11.4 mb).

Supplementary Table 1

– Follicle count. Total follicle and per patient counts in non-grafted controls, the 2-step/ASCs+OT group and the OT group. (PNG 62885 kb).

High resolution image (TIF 25749 kb).

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Cacciottola, L., Courtoy, G.E., Nguyen, T.Y.T. et al. Adipose tissue–derived stem cells protect the primordial follicle pool from both direct follicle death and abnormal activation after ovarian tissue transplantation. J Assist Reprod Genet 38, 151–161 (2021). https://doi.org/10.1007/s10815-020-02005-z

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