体外培养情况下猪卵母细胞脂质组形态功能状态参数分析

D. Starikova, T. Kuzmina
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引用次数: 0

摘要

基于细胞质内脂滴的形态和功能特征鉴定卵母细胞成熟的有效标记物将增加评估供体配子质量的标准的信息价值,从而提高在畜牧业、生物医学和兽医学中使用细胞生殖技术的效率。本研究的目的是分析体外培养前后susrofadesticus卵母细胞脂质组形态功能状态的参数(脂滴形态和尼罗红/脂滴复合物-尼罗红IF的荧光强度)。卵母细胞成熟使用Sage Media Cleavage (CooperSurgical, USA), 5%血清蛋白替代血清(CooperSurgical, USA), 10 IU人绒毛膜促性腺激素(Sigma, USA)。根据脂滴形态(颗粒状、簇状)和尼罗红IF(低、中、高)对供体卵母细胞群体进行排序,发现培养前尼罗红IF和LD平均呈颗粒状的卵母细胞比例为63%,显著高于尼罗红IF和LD平均呈混合状的配子比例41% (P<0.05)。培养后,中等尼罗红IF和颗粒状LD的卵母细胞水平为45%,超过相同尼罗红IF和混合LD的配子水平23% (P<0.05)。体外成熟卵母细胞具有高尼罗红IF和混合LD形态(颗粒和团簇)的比例为33%,培养前具有上述特征的比例为14% (P<0.05)。首次对susscrofadmesticus卵母细胞脂质体形态功能活性参数(形态学和IF尼罗红)之间的关系进行了比较形态学分析。揭示了培养前后脂质体的功能活性特征。利用脂质测定功能活性参数的综合分析评价卵母细胞的质量得到证实。猪卵母细胞的脂滴形态以颗粒形式表示,有希望用于培养,而尼罗河红IF被评估为低。对于所得结果的分析,建议在细胞生殖技术中采用提出的基于形态学和尼罗红IF的脂质组综合评估来评估供体卵母细胞的初始种群。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ANALYSIS OF PARAMETERS OF MORPHOFUNCTIONAL STATE OF PIG OOCYTE LIPIDOME IN CASE OF IN VITRO CULTIVATION
Identification of effective markers of oocyte maturation based on morphological and functional characteristics of intracytoplasmic lipid droplets will allow to increaseinformative value of the criteria for assessing the quality of donor gametes for improvement of the efficiency of using cellular reproductive technologies in animal husbandry, biomedicine, and veterinary medicine. The aim of the study is to analyze the parameters of morphofunctional state of the lipidome (the morphology of lipid droplets and the intensity of fluorescence of the Nile red/lipid droplets complex - Nile red IF) in Susscrofadomesticus oocytes before and after in vitro cultivation. Oocytes were matured using Sage Media Cleavage (CooperSurgical, USA) with 5% Serum Protein Substitute serum (CooperSurgical, USA), 10 IU human chorionic gonadotropin (Sigma, USA). As a result of ranking the population of donor oocytes according to morphology of lipid droplets (granules and clusters) and Nile red IF (low, medium, high), it was found that, the proportion of oocytes with an average Nile red IF and lipid droplets (LD) in the form of granules was 63% before cultivation, which was significantly higher than the proportion of gametes with an average IF of Nile red and LD of a mixed form of 41% (P<0.05). After cultivation, the level of oocytes with medium Nile red IF and LD in the form of granules was 45%, which exceeded the level of gametes with the same Nile red IF and mixed LD of 23% (P<0.05). The proportion of in vitro matured oocytes with high Nile red IF and mixed LD morphology (granules and clusters) was 33%, 14% were characterized by the above parameters before cultivation (P<0.05). Comparative morphological analysis of the relation between the parameters of morphofunctional activity (morphology and IF Nile red) of lipidome in Susscrofadomesticus oocytes was carried out for the first time. Features of functional activity of lipidome before and after cultivation were revealed. The usage of a comprehensive analysis of parameters of functional activity of lipidomeforassessment of the quality of the oocyte is substantiated. Pig oocytes which morphology of lipid droplets is represented by a granular form are promising for cultivation, while Nile red IF is assessed as low. With regard to the analysis of the obtained results, it is recommended to use the proposed comprehensive assessment of lipidome based on morphology and Nile red IF to assess the initial population of donor oocytes in cellular reproductive technologies.
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