Sebopela Maleke Dimpho, Mkhize Ntuthuko Raphael, Thema Mamonene Angelinah, Mphaphathi Masindi Lottus
Agricultural Research Council, Animal Production, Germplasm Conservation and Reproductive Biotechnologies, Private Bag X2, Pretoria 0062, South Africa.
School of Agricultural, Earth and Environmental Sciences, University of KwaZulu-Natal, Private Bag X01, Scottsville 3209, South Africa.
Vet Sci. 2025 May 12;12(5):461. doi: 10.3390/vetsci12050461.
This study aimed to compare the morphometry, morphology, and maturation capacity of cattle oocytes subjected to vitrification using different vitrification and maturation media. In Experiment 1, a total of 900 oocytes were divided into three groups: (1) matured before vitrification, (2) non-vitrified, and (3) vitrified as immature oocytes using the straw vitrification method. Morphometric parameters, including oocyte diameter, ooplasm, zona pellucida width (ZPW), granulosa cell width (GRSW), and zona pellucida-granulosa cell width (ZP GRSW), were measured (µm) before and after cryopreservation. In Experiment 2, the maturation capacity of three in vitro maturation (IVM) media (VitroMat-Protect™, BO-IVM™, and TCM199) was evaluated based on cumulus-oocyte complex (COC) expansion and polar body (PB) extrusion. Morphological abnormalities such as fragmented polar bodies (FPBs), large vacuoles (LVs), degenerated oocytes (DOs), and cracked cytoplasm (CC) were recorded. While vitrification did not significantly affect the oocyte diameter, ooplasm, or ZPW, it significantly reduced the GRSW and ZP GRSW. BO-IVM™ supported the highest COC expansion rate, while TCM199 had the lowest. Among vitrified oocytes, the highest PB extrusion rates were observed in BO-IVM (35.14 ± 5.01) and Vitromat-Protect (24.60 ± 5.67) as compared to TCM199 (18.44 ± 8.00; < 0.05). Oocytes with higher CC rates were observed in VitroMat-Protect™ (24.50 ± 10.53) and BO-IVM™ (31.42 ± 7.32) as compared to TCM199 (18.70 ± 7.04). In conclusion, the vitrification process affects the granulosa cells in both vitrified immature and mature oocytes. BO-IVM and VitroMat-Protect supported better oocyte maturation than TCM199, although vitrification increased FPB and CC rates.
本研究旨在比较使用不同玻璃化和成熟培养基进行玻璃化处理的牛卵母细胞的形态测量、形态学和成熟能力。在实验1中,总共900个卵母细胞被分为三组:(1)在玻璃化前成熟;(2)未玻璃化;(3)使用细管玻璃化法作为未成熟卵母细胞进行玻璃化。在冷冻保存前后测量形态测量参数,包括卵母细胞直径、卵质、透明带宽度(ZPW)、颗粒细胞宽度(GRSW)和透明带 - 颗粒细胞宽度(ZP GRSW)(μm)。在实验2中,基于卵丘 - 卵母细胞复合体(COC)扩展和极体(PB)排出评估三种体外成熟(IVM)培养基(VitroMat - Protect™、BO - IVM™和TCM199)的成熟能力。记录形态学异常,如破碎极体(FPB)、大液泡(LV)、退化卵母细胞(DO)和破裂细胞质(CC)。虽然玻璃化对卵母细胞直径、卵质或ZPW没有显著影响,但它显著降低了GRSW和ZP GRSW。BO - IVM™支持最高的COC扩展率,而TCM199最低。在玻璃化卵母细胞中,与TCM199(18.44 ± 8.00;P < 0.05)相比,在BO - IVM(35.14 ± 5.01)和Vitromat - Protect(24.60 ± 5.67)中观察到最高的PB排出率。与TCM199(18.70 ± 7.04)相比,在VitroMat - Protect™(24.50 ± 10.53)和BO - IVM™(31.42 ± 7.32)中观察到具有更高CC率的卵母细胞。总之,玻璃化过程影响玻璃化未成熟和成熟卵母细胞中的颗粒细胞。BO - IVM和VitroMat - Protect比TCM199支持更好的卵母细胞成熟,尽管玻璃化增加了FPB和CC率。