与卵巢实质细胞悬液共培养后小鼠腔前卵泡存活及发育的改善

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

作者信息

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

机构信息

Department of Basic Sciences and New Biological Technologies, Science and Culture University, Tehran, Iran.

Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.

出版信息

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

DOI:10.22074/ijfs.2023.1990372.1439

PMID:38368519

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10875308/

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.

摘要

背景

不孕症治疗和恶性肿瘤病例管理的同步持续改善，使生育力保存的潜力成为关注焦点。利用卵巢卵泡资源可有效提高生殖能力并预防不孕症。本研究的主要目的是尝试为小鼠卵泡生长创造适宜的体内环境。因此，探讨了卵巢实质细胞悬液对体外窦前卵泡生长和成熟的可能影响。

材料与方法

在本实验研究中，从12 - 14日龄NMRI小鼠的窦前卵泡中机械分离出卵巢实质细胞，然后分为5个实验组（G1：对照组，G2：新鲜卵泡与新鲜实质细胞悬液，G3：玻璃化冷冻 - 复温卵泡与新鲜实质细胞悬液，G4：新鲜卵泡与冻融实质细胞悬液，G5：玻璃化冷冻 - 复温卵泡与冻融实质细胞悬液）。在不同时间段检测卵泡和未成熟卵母细胞的直径、活力、腔形成、减数分裂恢复、体外受精（IVF）以及Gdf9、Bmp6和Bmp15基因表达。

结果

与对照组（G1：73.66%，G2：87.99%，G3：82.70%，G4：94.37%，G5：78.59%）相比，G2和G4组在第4天和第8天的卵泡和卵母细胞直径以及直至第12天的卵泡存活率显著更高。G3组和G5组生长标记基因的表达显著高于其他组，这表明实质细胞悬液对玻璃化溶液损伤的卵泡具有保护作用。