使用含低浓度 NaCl 的 αMEM 培养基进行成熟培养可提高猪体细胞核移植后卵母细胞的成熟和发育能力。

maturation using αMEM with reduced NaCl enhances maturation and developmental competence of pig oocytes after somatic cell nuclear transfer.

机构信息

College of Veterinary Medicine, Kangwon National University, Chuncheon 24341, Korea.

Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), College of Veterinary Medicine, Chungbuk National University and Institute of Stem Cell & Regenerative Medicine, Chungbuk National University, Cheongju 28644, Korea.

出版信息

J Vet Sci. 2022 Mar;23(2):e31. doi: 10.4142/jvs.21279.

DOI:10.4142/jvs.21279

PMID:35363440

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8977537/

Abstract

BACKGROUND

Compared to medium containing 108 mM sodium chloride (NaCl), maturation (IVM) using a simple medium with reduced (61.6 mM) NaCl increases the cytoplasmic maturation and embryonic development of pig oocytes.

OBJECTIVES

This study determines the effect of a complex medium containing reduced NaCl on the IVM and embryonic development of pig oocytes.

METHODS

Pig oocytes were matured in Minimum Essential Medium Eagle-alpha modification (αMEM) supplemented with 61.6 (61αMEM) or 108 (108αMEM) mM NaCl, and containing polyvinyl alcohol (PVA) (αMEMP) or pig follicular fluid (PFF) (αMEMF). Medium-199 (M199) served as the control for conventional IVM. Cumulus cell expansion, nuclear maturation, intra-oocyte glutathione (GSH) contents, size of perivitelline space (PVS), and embryonic development after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT) were evaluated after IVM.

RESULTS

Regardless of PVA or PFF supplementation, oocytes matured in 61αMEM showed increased intra-oocyte GSH contents and width of PVS ( < 0.05), as well as increased blastocyst formation ( < 0.05) after PA and SCNT, as compared to oocytes matured in 108αMEMP and M199. Under conditions of PFF-enriched αMEM, SCNT oocytes matured in 61αMEMF showed higher blastocyst formation ( < 0.05), compared to maturation in 108αMEMF and M199, whereas PA cultured oocytes showed no significant difference.

CONCLUSIONS

IVM in αMEM supplemented with reduced NaCl (61.6 mM) enhances the embryonic developmental competence subsequent to PA and SCNT, which attributes toward improved oocyte maturation.

摘要

背景

与含有 108 毫摩尔氯化钠（NaCl）的培养基相比，使用含有低浓度（61.6mM）NaCl 的简单培养基进行成熟（IVM）可提高猪卵母细胞的细胞质成熟和胚胎发育。

目的

本研究旨在确定含有低浓度 NaCl 的复杂培养基对猪卵母细胞 IVM 和胚胎发育的影响。

方法

猪卵母细胞在补充有 61.6（61αMEM）或 108（108αMEM）mM NaCl 的最小必需培养基 Eagle-alpha 修饰物（αMEM）中成熟，并含有聚乙烯醇（PVA）（αMEMP）或猪卵泡液（PFF）（αMEMF）。中 199（M199）作为常规 IVM 的对照。IVM 后评估卵丘细胞扩张、核成熟、卵内谷胱甘肽（GSH）含量、卵周隙（PVS）大小以及孤雌激活（PA）和体细胞核移植（SCNT）后的胚胎发育。

结果

无论是否添加 PVA 或 PFF，与在 108αMEMP 和 M199 中成熟的卵母细胞相比，在 61αMEM 中成熟的卵母细胞显示出更高的卵内 GSH 含量和 PVS 宽度（<0.05），以及更高的胚胎囊胚形成率（<0.05）PA 和 SCNT 后。在富含 PFF 的αMEM 条件下，与在 108αMEMF 和 M199 中成熟的卵母细胞相比，在 61αMEMF 中成熟的 SCNT 卵母细胞显示出更高的囊胚形成率（<0.05），而 PA 培养的卵母细胞则没有显著差异。