Comparison of CellRox green fluorescence upon thawing on in vitro Bos taurus and Bos indicus embryos cryopreserved by slow freezing or vitrification

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-09-18 DOI:10.1017/s0967199424000121

David Osornio, David Alejandro Contreras, Edgar Jimenez-Diaz, Tatiana Fiordelisio, Patricia López-Damian, José Francisco Martínez, Carlos Salvador Galina

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引用次数: 0

Abstract

Summary The aim of this study was to compare the levels of reactive oxygen species (ROS) in Bos taurus and Bos indicus in vitro embryos cryopreserved using either slow freezing or vitrification. Embryos were divided into four groups based on subspecies and freezing method: Bos indicus slow freezing (BiSF; n = 8), Bos indicus vitrification (BiVT; n = 10), Bos taurus slow freezing (BtSF; n = 9), and Bos taurus vitrification (BtVT; n = 6). After thawing, the embryos were incubated with CellRox Green and images were obtained using a confocal microscope. The fluorescence intensity of each cell was measured and expressed as arbitrary units of fluorescence (auf) and compared using a multiple regression and unpaired t-test with α = 0.05. Results showed that subspecies and the freezing method significantly affected auf (P < 0.001; R2 = 0.1213). Bos indicus embryos had higher auf than Bos taurus embryos, whether frozen by slow freezing (67.05 ± 23.18 vs 51.30 ± 16.84, P < 0.001) or vitrification (64.44 ± 23.32 vs 47.86 ± 17.53, P < 0.001). Slow freezing induced higher auf than vitrification in both Bos taurus (51.30 ± 16.84 vs 47.86 ± 17.53, P < 0.001) and Bos indicus (67.05 ± 23.18 vs 64.44 ± 23.32, P < 0.014). In conclusion, Bos taurus embryos had lower ROS levels when frozen using vitrification, while Bos indicus embryos had consistent ROS patterns regardless of the freezing method. However, Bos indicus embryos frozen by slow freezing tended to have a higher number of cells with elevated ROS levels.

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通过缓慢冷冻或玻璃化冷冻保存的体外牛胚胎和牛胚胎解冻时 CellRox 绿色荧光的比较

摘要本研究旨在比较采用缓慢冷冻或玻璃化技术冷冻保存的牛和籼体外胚胎中活性氧（ROS）的水平。胚胎根据亚种和冷冻方法分为四组：慢速冷冻组（BiSF；n = 8）、玻璃化组（BiVT；n = 10）、慢速冷冻组（BtSF；n = 9）和玻璃化组（BtVT；n = 6）。解冻后，用 CellRox Green 培养胚胎，并使用共聚焦显微镜获取图像。测量每个细胞的荧光强度，以任意荧光单位（auf）表示，并使用多元回归和非配对 t 检验（α = 0.05）进行比较。结果表明，亚种和冷冻方法对 auf 有显著影响（P < 0.001; R2 = 0.1213）。无论是慢速冷冻（67.05 ± 23.18 vs 51.30 ± 16.84，P < 0.001）还是玻璃化冷冻（64.44 ± 23.32 vs 47.86 ± 17.53，P < 0.001），Bos indicus 胚胎的 auf 均高于 Bos taurus 胚胎。在金牛（51.30 ± 16.84 vs 47.86 ± 17.53，P < 0.001）和麝香牛（67.05 ± 23.18 vs 64.44 ± 23.32，P < 0.014）中，慢速冷冻比玻璃化冷冻诱导更高的uf。总之，使用玻璃化技术冷冻的牛胚胎的 ROS 水平较低，而无论采用哪种冷冻方法，牛胚胎的 ROS 模式都是一致的。不过，用慢速冷冻法冷冻的牛胚胎往往有更多细胞的 ROS 水平升高。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.