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ID1 胞外多糖可改善玻璃化冷冻后体外生产牛胚胎的复苏效果。

Exopolysaccharide ID1 Improves Post-Warming Outcomes after Vitrification of In Vitro-Produced Bovine Embryos.

机构信息

Department of Animal Medicine and Surgery, Autonomous University of Barcelona, Cerdanyola del Vallès, ES-08193 Barcelona, Spain.

Brasuca In Vitro, Villahermosa MX-86040, Mexico.

出版信息

Int J Mol Sci. 2022 Jun 25;23(13):7069. doi: 10.3390/ijms23137069.

Abstract

This study aimed to assess the cryoprotectant role of exopolysaccharide (EPS) ID1, produced by Antarctic sp., in the vitrification of in vitro-produced (IVP) bovine embryos. IVP day 7 (D7) and day 8 (D8) expanded blastocysts derived from cow or calf oocytes were vitrified without supplementation (EPS0) or supplemented with 10 µg/mL (EPS10) or 100 µg/mL (EPS100) EPS ID1. The effect of EPS ID1 was assessed in post-warming re-expansion and hatching rates, differential cell count, apoptosis rate, and gene expression. EPS100 re-expansion rates were significantly higher than those observed for the EPS0 and EPS10 treatments, regardless of culture length or oocyte source. EPS100 hatching rate was similar to the one of the fresh blastocysts except for those D7 blastocysts derived from calf oocytes. No differences were observed among EPS ID1 treatments when the inner cell mass, trophectoderm, and total cell number were assessed. Although apoptosis rates were higher ( ≤ 0.05) in vitrified groups compared to fresh embryos, EPS100 blastocysts had a lower number ( ≤ 0.05) of apoptotic nuclei than the EPS0 or EPS10 groups. No differences in the expression of , , and genes between treatments were observed. Vitrification without EPS ID1 supplementation produced blastocysts with significantly higher gene expression, whereas treatment with 100 µg/mL EPS ID1 returned levels to those observed in non-vitrified blastocysts. Our results suggest that 100 µg/mL EPS ID1 added to the vitrification media is beneficial for embryo cryopreservation because it results in higher re-expansion and hatching ability and it positively modulates apoptosis.

摘要

本研究旨在评估南极 产生的胞外多糖 (EPS) ID1 在体外生产 (IVP) 牛胚胎玻璃化中的抗冻作用。对来自奶牛或小牛卵母细胞的 IVP 第 7 天 (D7) 和第 8 天 (D8) 扩张囊胚进行玻璃化处理,不添加(EPS0)或添加 10 µg/mL(EPS10)或 100 µg/mL(EPS100)EPS ID1。通过解冻后再扩张和孵化率、差异细胞计数、凋亡率和基因表达评估 EPS ID1 的作用。无论培养时间或卵母细胞来源如何,EPS100 的再扩张率均明显高于 EPS0 和 EPS10 处理组。EPS100 的孵化率与新鲜囊胚相似,但小牛卵母细胞来源的 D7 囊胚除外。当评估内细胞团、滋养外胚层和总细胞数时,EPS ID1 处理组之间没有差异。与新鲜胚胎相比,玻璃化组的凋亡率更高(≤0.05),但与 EPS0 或 EPS10 组相比,EPS100 囊胚的凋亡核数量更少(≤0.05)。在基因表达方面,未观察到各处理组之间 、 、 和 基因表达存在差异。不添加 EPS ID1 的玻璃化处理产生的囊胚中 基因表达显著升高,而添加 100 µg/mL EPS ID1 则使 水平恢复到非玻璃化囊胚的水平。我们的结果表明,在玻璃化培养基中添加 100 µg/mL EPS ID1 有利于胚胎的冷冻保存,因为它能提高再扩张和孵化能力,并能积极调节凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fd3e/9266775/aac31c46f02d/ijms-23-07069-g001.jpg

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