Jamalzaei Parisa, Rezazadeh Valojerdi Mojtaba, Montazeri Leila, Baharvand Hossein
Department of Pharmacology and Toxicology, School of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran.
Department of Anatomy, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Electronic Address:
Cell J. 2020 Jul;22(Suppl 1):49-60. doi: 10.22074/cellj.2020.6925. Epub 2020 Jul 18.
In the present study, the applicability of hyaluronic acid-alginate (HAA) hydrogel and ovarian cells (OCs) for the culture of mouse ovarian follicles were investigated and compared with those of alginate (ALG) and fibrin-alginate (FA) hydrogels.
In the first step of this experimental study, mechanically isolated preantral follicles from the ovaries of two-week-old mice were encapsulated in the absence or presence of OCs in ALG, HAA, and FA hydrogels and cultured for 14 days. The morphology, diameter, survival and antrum formation rates of the follicles and the maturation and quality of the oocytes were evaluated during culture. In the second step, preantral follicles were cultured similar to the first step, but for 13 days, and their gene expressions and hormonal secretion were assessed on the last day of culture.
In the absence of OCs, higher numbers of ALG- and HAA-encapsulated follicles reached the antral stage compared to FA-encapsulated follicles (P<0.05). However, a higher percentage of HAA-developed oocytes resumed meiosis up to the germinal vesicle breakdown (GVBD)/metaphase II (MII) stages in comparison with ALG-developed oocytes (P<0.05). HAA-encapsulated follicles had significant overexpression of most of the growth and differentiation genes, and secreted higher levels of estradiol (E2) compared to ALG- and FA-encapsulated follicles (P<0.05). The co-culture condition increased the diameter of ALG-encapsulated follicles on day 13 of culture (P<0.05). It also increased the survival and maturation rates of ALG- and FA-encapsulated follicles, respectively (P<0.05). The co-culture condition improved cortical granule distribution in all groups, increased E2 and progesterone (P4) secretions in the ALG and FA groups, and androstenedione (A4) secretion in the FA group (P<0.05).
The present study results show that HAA hydrogel is a promising hydrogel for follicle culture. OCs utilization could ameliorate the culture conditions regardless of the type of hydrogel.
在本研究中，研究了透明质酸 - 藻酸盐（HAA）水凝胶和卵巢细胞（OCs）用于小鼠卵巢卵泡培养的适用性，并与藻酸盐（ALG）和纤维蛋白 - 藻酸盐（FA）水凝胶进行比较。
在本实验研究的第一步中，将从两周龄小鼠卵巢中机械分离的窦前卵泡在不存在或存在OCs的情况下封装在ALG、HAA和FA水凝胶中，并培养14天。在培养过程中评估卵泡的形态、直径、存活率和窦腔形成率以及卵母细胞的成熟度和质量。在第二步中，窦前卵泡的培养方式与第一步相似，但培养13天，并在培养的最后一天评估其基因表达和激素分泌。
在不存在OCs的情况下，与FA封装的卵泡相比，ALG和HAA封装的卵泡达到窦腔阶段的数量更多（P<0.05）。然而，与ALG发育的卵母细胞相比，HAA发育的卵母细胞恢复减数分裂直至生发泡破裂（GVBD）/中期II（MII）阶段的百分比更高（P<0.05）。与ALG和FA封装的卵泡相比，HAA封装的卵泡大多数生长和分化基因有显著过表达，并且分泌更高水平的雌二醇（E2）（P<0.05）。共培养条件在培养第13天增加了ALG封装卵泡的直径（P<0.05）。它还分别提高了ALG和FA封装卵泡的存活率和成熟率（P<0.05）。共培养条件改善了所有组的皮质颗粒分布，增加了ALG和FA组中的E2和孕酮（P4）分泌以及FA组中的雄烯二酮（A4）分泌（P<0.05）。
本研究结果表明，HAA水凝胶是一种有前景的卵泡培养水凝胶。无论水凝胶类型如何，使用OCs都可以改善培养条件。
