Improvement of Mouse Preantral Follicle Survival and Development following Co-Culture with Ovarian Parenchyma Cell Suspension

Javad Najafi Salehi; Hussein Eimani; Abdolhossein Shahverdi; Mehdi Totonchi; Rouhollah Fathi; Seyed Akbar Moosavi; Seyed Mohamad Javad Taher Mofrad; Leila Sadat Tahaei

doi:10.22074/ijfs.2023.1990372.1439

Improvement of Mouse Preantral Follicle Survival and Development following Co-Culture with Ovarian Parenchyma Cell Suspension

Int J Fertil Steril. 2024 Feb 2;18(2):153-161. doi: 10.22074/ijfs.2023.1990372.1439.

Authors

Javad Najafi Salehi^{1

2}, Hussein Eimani³, Abdolhossein Shahverdi^{2

4}, Mehdi Totonchi⁵, Rouhollah Fathi², Seyed Akbar Moosavi⁶, Seyed Mohamad Javad Taher Mofrad², Leila Sadat Tahaei²

Affiliations

¹ Department of Basic Sciences and New Biological Technologies, Science and Culture University, Tehran, Iran.
² Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
³ Department of Basic Sciences and New Biological Technologies, Science and Culture University, Tehran, Iran.Email: eimanih@royaninstitute.org.
⁴ Reproductive Epidemiology Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
⁵ Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
⁶ Department of Lab Sciences, School of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.

Abstract

Background: The parallel and continued improvements in both infertility treatment and the management of malignancy cases have brought to the forefront the potential for fertility preservation. Using ovarian follicular resources can effectively improve reproductive capacity and prevent infertility. The primary aim of this research was to try to generate an appropriate in vivo environment for the growth of the mouse follicles. Hence, the possible effects of the ovarian parenchyma cell suspension were explored on the growth and maturation of preantral follicles in vitro.

Materials and methods: In this experimental study, ovarian parenchymal cells were mechanically dissociated from preantral follicles of 12-14 days-old NMRI mice and then divided into 5 experimental groups (G1: Control, G2: Fresh follicle with fresh parenchyma cell suspension, G3: Vitrified-warmed follicle with fresh parenchyma cell suspension, G4: Fresh follicle with frozen-thawed parenchyma cell suspension, and G5: Vitrified-warmed follicle with frozenthawed parenchyma cell suspension). The diameter of the follicles and immature oocytes, viability, antrum formation, resumption of meiosis, in vitro fertilization (IVF), and Gdf9, Bmp6, and Bmp15 gene expression were examined on different periods.

Results: The diameter of the follicles and the oocytes on days 4 and 8, as well as the survival rate of the follicles up to day 12, were significantly higher in G2 and G4 compared to the Ctrl group (G1: 73.66%, G2:87.99%, G3: 82.70%, G4: 94.37%, and G5: 78.59%). Expression of growth marker genes for G3, and G5 groups was significantly higher than other groups, which indicated the protective effects of parenchyma cell suspension on follicles damaged by vitrification solutions.

Conclusion: The growth, survival, and maturation of preantral follicles could be enhanced by co-culturing them with ovarian parenchyma cells. Further studies are needed to optimize the conditions for a successful parenchyma cell suspension-induced in vitro maturation (IVM) to occur in infertility clinics.

Keywords: Co-culture; Fertility Preservation; In Vitro Culture; Ovarian Tissue; Preantral Follicle.