Shams Mofarahe Zahra, Salehnia Mojdeh, Ghaffari Novin Marefat, Ghorbanmehr Nassim, Fesharaki Mohammad Gholami
Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Cell J. 2017 Apr-Jun;19(1):18-26. doi: 10.22074/cellj.2016.4890. Epub 2016 Dec 21.
This study was designed to evaluate the effects of vitrification and culture of human ovarian tissue on the expression of oocytic and follicular cell-related genes.
In this experimental study, ovarian tissue samples were obtained from eight transsexual women. Samples were cut into small fragments and were then assigned to vitrified and non-vitrified groups. In each group, some tissue fragments were divided into un-cultured and cultured (in α-MEM medium for 2 weeks) subgroups. The normality of follicles was assessed by morphological observation under a light microscope using hematoxylin and eosin (H&E) staining. Expression levels of factor in the germ line alpha (), KIT ligand (), growth differentiation factor 9 () and follicle stimulating hormone receptor () genes were quantified in both groups by real-time reverse transcriptase polymerase chain reaction (RT-PCR) at the beginning and the end of culture.
The percentage of normal follicles was similar between non-cultured vitrified and non-vitrified groups (P>0.05), however, cultured tissues had significantly fewer normal follicles than non-cultured tissues in both vitrified and non-vitrified groups (P<0.05). In both cultured groups the rate of primary and secondary follicles was significantly higher than non-cultured tissues (P<0.05). The expression of all examined genes was not significantly altered in both non-cultured groups. Whiles, in comparison with cultured tissues non-cultured tissues, the expression of gene was significantly decreased, gene was not changed, and genes was significantly increased (P<0.05).
Human ovarian vitrification following culture has no impairing effects on follicle normality and development and expression of related-genes. However, culture condition has deleterious effects on normality of follicles.
本研究旨在评估人卵巢组织玻璃化冷冻及培养对卵母细胞和卵泡细胞相关基因表达的影响。
在本实验研究中,从八名变性女性获取卵巢组织样本。将样本切成小碎片,然后分为玻璃化冷冻组和非玻璃化冷冻组。每组中,一些组织碎片再分为未培养组和培养组(在α - MEM培养基中培养2周)。使用苏木精和伊红(H&E)染色,通过光学显微镜下的形态学观察评估卵泡的正常性。在培养开始和结束时,通过实时逆转录聚合酶链反应(RT-PCR)对两组中生殖系α()、KIT配体()、生长分化因子9()和卵泡刺激素受体()基因的表达水平进行定量分析。
未培养的玻璃化冷冻组和非玻璃化冷冻组之间正常卵泡的百分比相似(P>0.05),然而,在玻璃化冷冻组和非玻璃化冷冻组中,培养后的组织正常卵泡均明显少于未培养的组织(P<0.05)。在两个培养组中,初级和次级卵泡的比例均显著高于未培养的组织(P<0.05)。在两个未培养组中,所有检测基因的表达均无显著变化。同时,与培养后的组织相比,未培养的组织中基因表达显著降低,基因无变化,基因和基因显著增加(P<0.05)。
培养后人卵巢玻璃化冷冻对卵泡正常性、发育及相关基因表达无损害作用。然而,培养条件对卵泡正常性有有害影响。