Hashemian Zohreh, Afsharian Parvaneh, Farzaneh Parvaneh, Eftekhari-Yazdi Poopak, Vakhshiteh Faezeh, Daneshvar Amoli Abdolreza, Nasimian Ahmad
Human and Animal Cell Bank, Iranian Biological Resource Center (IBRC), ACECR, Tehran, Iran.
Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Cytotechnology. 2020 Sep 28;72(6):833-45. doi: 10.1007/s10616-020-00426-3.
Oocyte maturation is an important phase in fertility and any disorder in this process could lead to infertility. The most common disorder during folliculogenesis is polycystic ovary syndrome (PCOS). Due to the secretive activity of granulosa cells (GCs), they play a vital role in folliculogenesis. Although scientists use various cellular and molecular methods to have a better understanding of the mechanism of these cells, some limitations still exist in GC culture such as low primary cell yield and proliferation capability. Therefore, immortalization of primary cells is an approach to overcome these limitations. In the current study, GCs were obtained from two females, one with PCOS and one with normal folliculogenesis. In the first stage, we established two human GC (hGC) lines by immortalizing them through retrovirus-mediated transfer of the human telomerase reverse transcriptase (hTERT) and c-Myc genes. Subsequently, the normal and PCOS cell lines were characterized and were investigated for their growth features. The cell lines were also examined in terms of immortal markers of hTERT, follicle stimulating hormone receptor (FSHR), aromatase, anti-Müllerian hormone (AMH), growth differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), estrogen, and progesterone. Our results indicated that the normal and PCOS cell lines both showed similar characteristics to GCs during the follicular stage in normal and PCOS women. The normal and PCOS cell lines demonstrate molecular mechanisms similar to that of GCs such as folliculogenesis, oogenesis, and steroidogenesis, which enable researchers to perform further investigations in future.
卵母细胞成熟是生育过程中的一个重要阶段,这一过程中的任何紊乱都可能导致不孕。卵泡发生过程中最常见的紊乱是多囊卵巢综合征(PCOS)。由于颗粒细胞(GCs)的分泌活动,它们在卵泡发生中起着至关重要的作用。尽管科学家们使用各种细胞和分子方法来更好地了解这些细胞的机制,但GC培养仍存在一些局限性,如原代细胞产量低和增殖能力有限。因此,原代细胞永生化是克服这些局限性的一种方法。在本研究中,从两名女性身上获取了颗粒细胞,一名患有PCOS,另一名卵泡发生正常。在第一阶段,我们通过逆转录病毒介导的人类端粒酶逆转录酶(hTERT)和c-Myc基因转移使颗粒细胞永生化,从而建立了两个人类颗粒细胞(hGC)系。随后,对正常和PCOS细胞系进行了表征,并研究了它们的生长特性。还从hTERT、促卵泡激素受体(FSHR)、芳香化酶、抗苗勒管激素(AMH)、生长分化因子9(GDF9)、骨形态发生蛋白15(BMP15)、雌激素和孕酮的永生化标志物方面对细胞系进行了检测。我们的结果表明,正常和PCOS细胞系在正常和PCOS女性的卵泡期均表现出与颗粒细胞相似的特征。正常和PCOS细胞系表现出与颗粒细胞相似的分子机制,如卵泡发生、卵子发生和类固醇生成,这使研究人员能够在未来进行进一步的研究。