Tanbakooei Sara, Haramshahi Seyed Mohammad Amin, Vahabzadeh Gelareh, Barati Mahmood, Katebi Majid, Golab Fereshteh, Shetabi Qazal, Niknam Narges, Roudbari Leila, Rajabi Fomeshi Motahareh, Amini Moghadam Soheila
Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, Iran.
Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
J Reprod Infertil. 2021 Oct-Dec;22(4):241-250. doi: 10.18502/jri.v22i4.7649.
obtaining oocytes can be an appropriate alternative for patients with gonadal insufficiency or cancer survivors. The purpose of the current research was isolating stem cells from ovarian cortical tissue as well as evaluating the effectiveness of follicle stimulating hormone (FSH), basic fibroblast growth factor (bFGF), and neurotrophin 3 (NT3) in differentiating to oocyte-like cells.
A human ovary was dissected and cortical tissue pieces were cultured for cell isolation. Isolated cells were divided into 8 groups (3 cases in each group) of control, FSH, NT3, bFGF, FSH+NT3, FSH+bFGF, NT3+bFGF, and FSH+NT3+ bFGF. Pluripotency specific gene (OCT4-A and Nanog), initial germ cells (c-KIT and VASA) and PF growth initiators (GDF-9 and Lhx-8) were evaluated by qRTPCR. Experiments were performed in triplicate and there were 3 samples in each group. The results were analyzed using one-way ANOVA and p-value less than 0.05 was considered statistically significant.
Flow cytometry results showed that cells isolated from the ovarian cortex expressed markers of pluripotency. The results showed that the expression of Nanog, OCT4, GDF-9 and VASA was significantly increased in FSH+NT3 group, while treatment with bFGF caused significant expression of c-KIT and Lhx-8 (p<0.05). Also, according to the results, isolated cells treated with NT3 significantly increased c-KIT expression.
According to our results, the ovarian cortex cells could be differentiated into primordial follicles if treated with the proper combination of FSH, bFGF, and NT3. These findings provided a new perspective for the future of gamete proudest.
对于性腺功能不全患者或癌症幸存者而言，获取卵母细胞可能是一种合适的替代方案。当前研究的目的是从卵巢皮质组织中分离干细胞，并评估卵泡刺激素（FSH）、碱性成纤维细胞生长因子（bFGF）和神经营养因子3（NT3）在分化为卵母细胞样细胞方面的有效性。
解剖人卵巢并培养皮质组织块以进行细胞分离。将分离出的细胞分为8组（每组3例），分别为对照组、FSH组、NT3组、bFGF组、FSH + NT3组、FSH + bFGF组、NT3 + bFGF组和FSH + NT3 + bFGF组。通过qRTPCR评估多能性特异性基因（OCT4 - A和Nanog）、原始生殖细胞（c - KIT和VASA）以及卵泡生长启动因子（GDF - 9和Lhx - 8）。实验重复进行3次，每组有3个样本。结果采用单因素方差分析进行分析，p值小于0.05被认为具有统计学意义。
流式细胞术结果显示，从卵巢皮质分离的细胞表达多能性标志物。结果表明，FSH + NT3组中Nanog、OCT4、GDF - 9和VASA的表达显著增加，而用bFGF处理导致c - KIT和Lhx - 8的显著表达（p < 0.05）。此外，根据结果，用NT3处理的分离细胞显著增加了c - KIT的表达。
根据我们的结果，如果用FSH、bFGF和NT3的适当组合处理，卵巢皮质细胞可分化为原始卵泡。这些发现为配子产生的未来提供了新的视角。
