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由于线粒体膜电位和活性氧水平的改变,用透明质酸培养的猪胚胎质量得到改善。

Improved quality of porcine embryos cultured with hyaluronan due to the modification of the mitochondrial membrane potential and reactive oxygen species level.

作者信息

Romek Marek, Gajda Barbara, Krzysztofowicz Ewa, Kucia Marcin, Uzarowska Agnieszka, Smorag Zdzislaw

机构信息

Department of Cell Biology and Imaging, Institute of Zoology, Jagiellonian University, Gronostajowa 9, 30-387 Krakow, Poland.

Department of Animal Reproduction Biotechnology, National Research Institute of Animal Production, 32-083 Balice/Krakow, Poland.

出版信息

Theriogenology. 2017 Oct 15;102:1-9. doi: 10.1016/j.theriogenology.2017.06.026. Epub 2017 Jun 29.

Abstract

Although considerable progress has been made in pig embryo culture systems, the developmental competence and quality of the produced embryos are still lower than their in vivo-derived counterparts. Because hyaluronan (HA) regulates various cellular processes and possesses antioxidant properties, this glycosaminoglycan seems to be a promising supplement in culture media. However, until now, its beneficial influence on in vitro pig embryo development has been debatable. Hence, we aimed to investigate the effect of 0.25 mg/mL, 0.5 mg/mL and 1 mg/mL concentrations of HA on the developmental potential and quality of cultured porcine embryos. We found that 1 mg/mL HA supplementation significantly increased the obtained percentages of cleaved embryos to ∼95%, morulae to ∼87% and blastocysts to ∼77%. At 0.5 mg/mL and 1 mg/mL HA concentrations, we observed a significantly improved blastocyst quality, expressed as the total number of cells per blastocyst, number of cells in the inner cell mass, number of TUNEL-positive nuclei per blastocyst, the TUNEL index and the blastocyst diameter. Because the inner mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) level are important for proper embryo development, for the first time, we measured these two parameters in cultured embryos at various HA concentrations and during their development up to the expanded blastocyst stage. For blastocysts cultured with 1 mg/mL HA, the ΔΨm and ROS level were ∼1.6 and 2.7 times lower, respectively, than those of the control blastocysts. Both ΔΨm and the ROS level were increased in parallel during in vitro embryo development with and without HA, but this increase was less pronounced in the presence of HA. Hence, our quantitative data unequivocally show that supplementation of NCSU-23 culture medium with 1 mg/mL HA improves the developmental potential and quality of pig embryos. This effect results from a significant decrease in the ROS level induced by the HA-dependent ΔΨm reduction.

摘要

尽管猪胚胎培养系统已取得了显著进展,但所产生胚胎的发育能力和质量仍低于体内来源的胚胎。由于透明质酸(HA)调节多种细胞过程并具有抗氧化特性,这种糖胺聚糖似乎是培养基中一种有前景的补充剂。然而,到目前为止,其对体外猪胚胎发育的有益影响一直存在争议。因此,我们旨在研究0.25mg/mL、0.5mg/mL和1mg/mL浓度的HA对培养的猪胚胎发育潜力和质量的影响。我们发现,添加1mg/mL的HA可使分裂胚的获得率显著提高至约95%,桑葚胚提高至约87%,囊胚提高至约77%。在HA浓度为0.5mg/mL和1mg/mL时,我们观察到囊胚质量显著提高,表现为每个囊胚的细胞总数、内细胞团中的细胞数、每个囊胚中TUNEL阳性细胞核的数量、TUNEL指数和囊胚直径。由于线粒体内膜电位(ΔΨm)和活性氧(ROS)水平对胚胎的正常发育很重要,我们首次在不同HA浓度的培养胚胎及其发育至扩张囊胚阶段的过程中测量了这两个参数。对于用1mg/mL HA培养的囊胚,其ΔΨm和ROS水平分别比对照囊胚低约1.6倍和2.7倍。在有或没有HA的体外胚胎发育过程中,ΔΨm和ROS水平均同时升高,但在有HA的情况下这种升高不太明显。因此,我们的定量数据明确表明,在NCSU-23培养基中添加1mg/mL的HA可提高猪胚胎的发育潜力和质量。这种作用源于HA依赖性ΔΨm降低所诱导的ROS水平显著降低。

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