模拟生理卵母细胞成熟(SPOM)和左旋肉碱对牛卵母细胞发育能力的影响

Q3 Biochemistry, Genetics and Molecular Biology Avicenna journal of medical biotechnology Pub Date : 2024-10-01 DOI:10.18502/ajmb.v16i4.16742
Ali Malekpour, Abolfazl Shirazi, Sara Borjian Boroujeni, Ali Sarvari, Mohammad Mehdi Naderi, Mostafa Pournourali, Bahareh Behzadi, Mohammad Mehdi Mehrazar
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引用次数: 0

摘要

背景:模拟卵母细胞生理成熟(SPOM)在体外模拟了卵母细胞在cAMP调节剂作用下成熟的生理过程。这些调节剂可增加细胞内的 cAMP 浓度,从而抑制减数分裂的立即恢复,使卵母细胞有更多时间获得最佳发育能力。此外,左旋肉碱还有助于通过脂肪酸的β-氧化作用增加细胞的能量供应。本研究旨在探讨在体外成熟(IVM)和体外培养(IVC)期间补充SPOM和左旋肉碱对牛卵母细胞发育能力的影响:在体外成熟(IVM)或体外培养(IVC)的前2小时(pre-IVM)有或无福斯可林+IBMX、有或无左旋肉碱(LC)的情况下培养卵母细胞复合体(COCs),分为以下6个实验组:I) IVM 前(IVM 前组);II) IVM 前,IVM 期间补充左旋肉碱(IVM 前/LC 组);III) IVM 期间补充左旋肉碱(IVM/LC 组);IV) IVM 期间补充左旋肉碱(IVM/LC 组);IV) IVM 期间补充左旋肉碱(IVM/LC 组)。体外培养期间补充左旋肉碱组(IVC/LC 组),V) IVM 前+ IVC/LC 组,VI) IVM 和 IVC 期间不处理组(对照组)。评估囊胚的分裂率和囊胚率、囊胚总细胞数以及囊胚中 Nanog、Bax、Oct4、Cdx2 和 Ifnt 基因的表达。为评估实验组之间的差异,首先采用单因素方差分析,然后进行事后费舍尔 LSD 分析。当 pResults 达到统计学意义时,认为组间差异显著:除 IVC/LC 组和 IVM/LC 组外,Pre-IVM 组和 Pre-IVM/LC 组的卵裂率和囊胚率分别高于对照组和其他组(p≤0.05)。IVM前组和IVM前/LC组囊胚内细胞团(ICM)的数量以及ICM/TE的比率均高于对照组(p结论:SPOM的应用对囊胚内细胞团的数量以及ICM/TE的比率均有影响:总之,在牛卵母细胞体外受精过程中应用 SPOM 适应性体外受精和左旋肉碱可提高胚胎的数量和质量。
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The Effect of Simulated Physiological Oocyte Maturation (SPOM) and L-Carnitine on Bovine Oocyte Developmental Competence.

Background: Simulated Physiological Oocyte Maturation (SPOM) mimics in vitro the physiological events of oocyte maturation in the presence of cAMP modulators. These modulators increase the intracellular concentrations of cAMP, which inhibits the immediate resumption of meiosis and gives the oocyte more time to gain optimal developmental competence. In addition, L-carnitine helps to increase the energy supply of cells through the β-oxidation of fatty acids. This study aimed to investigate the effect of SPOM and L-carnitine supplementation during In Vitro Maturation (IVM) and In Vitro Culture (IVC) on the developmental competence of bovine oocytes.

Methods: Ovarian Cumulus Complexes (COCs) were cultured in the presence or absence of forskolin+IBMX during the first 2 hr of IVM (pre-IVM) with or without L-carnitine (LC) during IVM or IVC in six experimental groups as follows: I) pre-IVM (pre-IVM group), II) pre-IVM with L-carnitine supplementation during IVM (pre-IVM/LC group), III) L-carnitine supplementation during IVM (IVM/LC group), IV) L-carnitine supplementation during in vitro culture (IVC/LC group), V) pre-IVM+ IVC/LC group, and VI) no treatment during IVM and IVC (Control group). The cleavage and blastocyst rates, the blastocysts' total cells number, and the expression of Nanog, Bax, Oct4, Cdx2, and Ifnt genes in resulting blastocysts were assessed. To assess differences among experimental groups, a one-way analysis of variance was initially employed, followed by post hoc Fisher LSD. The difference between groups was considered statistically significant when p<0.05.

Results: The cleavage and blastocyst rates in the Pre-IVM and Pre-IVM/LC groups was higher than control group and other groups (p≤0.05) except for IVC/LC and IVM/LC groups, respectively. The number of blastocyst's Inner Cell Mass (ICM) in pre-IVM and Pre-IVM/LC groups as well as the ratio of ICM/TE were higher than control group (p<0.05). The expression of OCT4, CDX2, and IFNT increased in both the pre-IVM and pre-IVM/LC groups compared to the control group (p<0.05).

Conclusion: In conclusion, the application of SPOM-adapted IVM and L-carnitine during IVM of bovine oocyte improves the quantity and quality of the resulting embryos.

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来源期刊
Avicenna journal of medical biotechnology
Avicenna journal of medical biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
2.90
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43
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