Castañeda Olga Juliana Roldan, de Aguiar Francisco Léo Nascimento, de Sá Naiza Arcângela Ribeiro, Morais Maria Luana Galdencio Dos Santos, Cibin Francielli Weber Santos, Torres Ciro Alexandre Alves, de Figueiredo José Ricardo
Universidade Estadual do Ceará, Laboratório de Manipulação de Oócitos Inclusos em Folículos Ovarianos Pré-antrais, Fortaleza, CE, Brasil.
Universidade Federal de Viçosa, Laboratório de Fisiologia Animal e Reprodução, Viçosa, MG, Brasil.
Anim Reprod. 2019 Nov 18;16(4):838-845. doi: 10.21451/1984-3143-AR2019-0011.
This study evaluated a powdered coconut water solution (ACP 406®) as a base culture medium on the survival and development of goat preantral follicles. The ovarian fragments were either immediately fixed in Carnoy solution (non-cultured control) or individually cultured for 2 or 6 days. The following culture media (all containing 100 μg/mL penicillin and 100 μg/mL streptomycin) were evaluated: α-MEM (α-MEM alone, without additional supplementation); α-MEM+ (supplemented α-MEM); ACP (ACP®406 alone); or ACP+ (supplemented ACP®406). Additional supplementation includes: 1.25 mg/mL bovine serum albumin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 2 mM glutamine, and 2 mM hypoxanthine. The endpoints (i) follicular morphology; (ii) development; (iii) estradiol production; and (iv) reactive oxygen species (ROS) were recorded. Data were analyzed using chi-square, Turkey, t-test or One-Way ANOVA. Differences were considered significant when P < 0.05. At day 2 of culture, a greater (P < 0.05) percentage of morphologically normal follicles was observed between ACP+ and ACP treatments. Moreover, at day 2 of culture, no hormonal difference (P < 0.05) was observed between ACP+ and both α-MEM treatments. At day 6 of culture when ACP and α-MEM treatments were compared the percentage of healthy follicles were similar (P > 0.05) among treatments. Overall, all treatments had lower primordial follicles (P < 0.05) accompany by greater developing follicles (P < 0.05) percentages than non-cultured control treatment, indicating primordial follicle activation. However, at day 6 of culture, the percentage of primordial follicle development were similar (P > 0.05) among the treatments. Likewise, no differences (P > 0.05) were observed for ROS production and follicular and oocyte diameters among treatments. Therefore, ACP+ has the equivalent efficiency to MEM+ in maintaining the survival and development of goat preantral follicles, representing an alternative plant-based low-cost culture medium for culture.
本研究评估了一种椰子水粉末溶液（ACP 406®）作为基础培养基对山羊腔前卵泡存活和发育的影响。卵巢组织块要么立即固定于卡诺伊溶液中（非培养对照组），要么单独培养2天或6天。评估了以下培养基（均含有100μg/mL青霉素和100μg/mL链霉素）：α-MEM（仅α-MEM，无额外添加物）；α-MEM+（添加后的α-MEM）；ACP（仅ACP®406）；或ACP+（添加后的ACP®406）。额外添加物包括：1.25mg/mL牛血清白蛋白、10μg/mL胰岛素、5.5μg/mL转铁蛋白、5ng/mL硒、2mM谷氨酰胺和2mM次黄嘌呤。记录了以下终点指标：（i）卵泡形态；（ii）发育情况；（iii）雌二醇产生量；以及（iv）活性氧（ROS）。数据采用卡方检验、土耳其检验、t检验或单因素方差分析进行分析。当P<0.05时，差异被认为具有统计学意义。在培养第2天，ACP+组和ACP组之间观察到形态正常卵泡的比例更高（P<0.05）。此外，在培养第2天，ACP+组与两种α-MEM组之间未观察到激素差异（P<0.05）。在培养第6天，比较ACP组和α-MEM组时，各处理组中健康卵泡的比例相似（P>0.05）。总体而言，与非培养对照组相比，所有处理组的原始卵泡比例均较低（P<0.05），而发育中卵泡的比例均较高（P<0.05），表明原始卵泡被激活。然而，在培养第6天，各处理组中原始卵泡发育的比例相似（P>0.05）。同样，各处理组在ROS产生量以及卵泡和卵母细胞直径方面未观察到差异（P>0.05）。因此，ACP+在维持山羊腔前卵泡存活和发育方面具有与MEM+相当的效率，是一种可供选择的低成本植物源培养基用于培养。
