Impact of cholesterol supplementation on Pantaneiro bovine semen cryopreservation: Insights into in vitro embryo production

IF 2.4 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology Pub Date : 2024-12-18 DOI:10.1016/j.theriogenology.2024.12.014

Fernanda Guadagnin , Luciana Keiko Hatamoto-Zervoudakis , Walter Augusto dos Santos Marinho , Cassia Panizzon Dal Curtivo , Thaiza Thommen Maciel Preti , Raphael Campos Quinteiro , Tathiana Ferguson Motheo

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Abstract

This study aimed to evaluate the impact of cholesterol supplementation at various concentrations in cryopreserved Pantaneiro bovine semen on in vitro embryo production (IVEP). Grade I and II cumulus-oocyte complexes (COCs) were collected from ovaries retrieved from a commercial slaughterhouse and matured in vitro for 24 h. The matured COCs were divided into four groups based on the concentration of cholesterol -loaded cyclodextrin (CLC) during semen cryopreservation from a Pantaneiro breed bull: Control (C) - 0 mg/mL CLC, T1 - 0.5 CLC, T2 - 1 mg/mL CLC, and T3 - 1.5 mg/mL CLC. After 18 h of incubation, structures were denuded and transferred to in vitro culture (CIV) for 8 days. Cleavage rate was assessed on the second day (D2), and on the seventh day (D7), embryo classification and blastocyst production rate were evaluated. Additionally, total and apoptotic embryonic cell counts were conducted using differential staining for bovine embryos. Oxidative status was assessed in the FIV and CIV media of each treatment by measuring thiobarbituric acid reactive substances – TBARS. Concentrations of 1.0 mg/mL and 1.5 mg/mL of CLC significantly reduced the proportion of cleaved structures (P = 0.0029), as well as the percentages of cleaved structures and blastocysts (P < 0.001). Moreover, increasing CLC concentrations decreased the total number of embryonic cells (P < 0.001). No significant differences were noticed in other parameters. In conclusion, supplementing cryopreserved Pantaneiro cattle semen with cholesterol-loaded cyclodextrin (CLC) at concentrations of 0.5, 1, and 1.5 mg/mL adversely affects in vitro embryo production.

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补充胆固醇对潘塔内罗牛精液冷冻保存的影响：对体外胚胎生产的见解。

本研究旨在评价在低温保存的潘塔内罗牛精液中添加不同浓度的胆固醇对体外胚胎生成（IVEP）的影响。从商业屠宰场的卵巢中收集I级和II级卵母细胞复合物（COCs），并在体外成熟24 h。根据潘塔内罗种公牛精液冷冻保存过程中胆固醇负载环糊精（CLC）的浓度将成熟的COCs分为4组：对照(C) - 0 mg/mL CLC、T1 - 0.5 mg/mL CLC、T2 - 1 mg/mL CLC和T3 - 1.5 mg/mL CLC。孵育18 h后，剥去结构，转入体外培养（CIV） 8天。在第2天（D2）评估卵裂率，第7天（D7）评估胚胎分类和囊胚产生率。此外，用差异染色法对牛胚胎进行总胚胎细胞计数和凋亡细胞计数。通过测量硫代巴比妥酸反应物质- TBARS来评估每个处理的FIV和CIV培养基中的氧化状态。1.0 mg/mL和1.5 mg/mL的CLC浓度显著降低了裂裂结构的比例（P = 0.0029），并显著降低了裂裂结构和囊胚的比例（P = 0.0029）

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来源期刊

Theriogenology 农林科学-生殖生物学

CiteScore

5.50

自引率

14.30%

发文量

387

审稿时长

72 days

期刊介绍： Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.