海藻酸钠浓度和卵巢细胞对小鼠腔前卵泡体外发育的影响：析因研究
Effects of Alginate Concentration and Ovarian Cells on In Vitro Development of Mouse Preantral Follicles: A Factorial Study.
作者信息
Jamalzaei Parisa, Valojerdi Mojtaba Rezazadeh, Montazeri Leila, Baharvand Hossein
机构信息
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
出版信息
Int J Fertil Steril. 2020 Jan;13(4):330-338. doi: 10.22074/ijfs.2020.5746. Epub 2019 Nov 11.
BACKGROUND
In the present study, the effects of alginate (ALG) concentration and ovarian cells (OCs) on the development and function of follicles were simultaneously evaluated.
MATERIALS AND METHODS
In the first step of this experimental study, preantral follicles were isolated from the ovaries of 2-week-old mice, encapsulated in the absence or presence of OCs in 0.5, 0.75 and 1% ALG hydrogels, and cultured for 14 days. The morphology, diameter, survival and antrum formation rates of the follicles and the maturation of the oocytes were evaluated during culture. In the second step, preantral follicles were cultured in the best chosen ALG concentration, in both the absence and presence of OCs. Following these steps, the amount of DNA fragmentation, the expression levels of connexin 37 and connexin 43 proteins, the secretion levels of estradiol, progesterone and androstenedione by the follicles and the quality of mature (MII) oocytes were assessed.
RESULTS
Our data revealed that in the absence of OCs, follicles of 0.5% group showed a higher survival rate than the 0.75 and 1% groups (71.87 vs. 52.52 and 40%, respectively, P<0.05). Nonetheless, the antrum formation rate of the 1% group was higher and its oocyte degeneration rate was lower than that in the other groups. Furthermore, it was observed that co-culture of follicles with OCs relatively increased the follicle diameter, survival, antrum formation, and germinal vesicle (GV) to GV break down (GVBD)/MII transition rates. At last, the comparison of 0.5%-OCs and 0.5%+OCs groups indicated that the co-culture condition resulted in more progesterone production (1.8 ± 0.2 vs. 3.2 ± 0.4 ng/ml, respectively, P<0.05) and also decreased oocytes' cortical granule abnormalities (100 vs. 40% for 0.5%- OCs and 0.5%+OCs groups, respectively).
CONCLUSION
The present study revealed that 0.5% ALG hydrogel is relatively suitable for preantral follicle culture, and in the presence of OCs, it mimics the natural ovarian condition better than the higher concentrations of ALG hydrogel.
背景
在本研究中，同时评估了藻酸盐（ALG）浓度和卵巢细胞（OCs）对卵泡发育和功能的影响。
材料与方法
在本实验研究的第一步中，从2周龄小鼠的卵巢中分离出窦前卵泡，在不存在或存在OCs的情况下，将其包裹在0.5%、0.75%和1%的ALG水凝胶中，并培养14天。在培养过程中评估卵泡的形态、直径、存活率和腔形成率以及卵母细胞的成熟情况。在第二步中，将窦前卵泡在最佳选择的ALG浓度下进行培养，分别在不存在和存在OCs的情况下培养。经过这些步骤后，评估DNA片段化的量、连接蛋白37和连接蛋白43蛋白的表达水平、卵泡分泌雌二醇、孕酮和雄烯二酮的水平以及成熟（MII）卵母细胞的质量。
结果
我们的数据显示，在不存在OCs的情况下，0.5%组的卵泡存活率高于0.75%和1%组（分别为71.87% vs. 52.52%和40%，P<0.05）。然而，1%组的腔形成率更高，其卵母细胞退化率低于其他组。此外，观察到卵泡与OCs共培养相对增加了卵泡直径、存活率、腔形成以及生发泡（GV）到生发泡破裂（GVBD）/MII转变率。最后，0.5%-OCs组和0.5%+OCs组的比较表明，共培养条件导致更多的孕酮产生（分别为1.8±0.2 vs. 3.2±0.4 ng/ml，P<0.05），并且还降低了卵母细胞的皮质颗粒异常（0.5%-OCs组和0.5%+OCs组分别为100% vs. 40%）。
结论
本研究表明，0.5%的ALG水凝胶相对适合窦前卵泡培养，并且在存在OCs的情况下，它比更高浓度的ALG水凝胶更能模拟自然卵巢条件。
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