Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

In vitro oocyte maturation in a medium containing reduced sodium chloride improves the developmental competence of pig oocytes after parthenogenesis and somatic cell nuclear transfer

Joohyeong Lee A B , Hanna Lee A , Yongjin Lee A , Bola Park A , Fazle Elahi A , Seung Tae Lee C , Choon-Keun Park C , Sang-Hwan Hyun D and Eunsong Lee A B E

A Laboratory of Theriogenology, College of Veterinary Medicine, Kangwon National University, Chuncheon, Gangwon 24341, Korea.

B Institute of Veterinary Science, Kangwon National University, Chuncheon, Gangwon 24341, Korea.

C Division of Applied Animal Science, College of Animal Life Science, Kangwon National University, Chuncheon, Gangwon 24341, Korea.

D Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 28644, Korea.

E Corresponding author. Email: eslee@kangwon.ac.kr

Reproduction, Fertility and Development - https://doi.org/10.1071/RD15488
Submitted: 24 November 2015  Accepted: 3 August 2016   Published online: 5 September 2016

Abstract

The present study investigated the effects of IVM in hypotonic medium containing reduced (61.6 mM) NaCl compared with isotonic medium containing 108.0 mM NaCl (designated L and N respectively) on oocyte maturation and embryonic development in pigs. IVM culture was divided into four periods at 11-h intervals. Oocytes cultured in N for 33 h and then in L for 11 h of IVM (N-N-N-L) showed significantly improved (P < 0.05) nuclear maturation of oocytes (75.4–79.0% vs 60.2–85.8%) and blastocyst formation (61.5–66.1% vs 45.2–67.5%) after parthenogenesis (PA) compared with other treatments (L-L-L-L, L-L-L-N, L-L-N-L, N-N-L-L, N-N-L-N, L-L-N-L, L-N-N-L and N-L-N-L). Oocytes matured in L-L-L-L and N-N-N-L had an increased (P < 0.05) perivitelline space (11.0–12.5 vs 5.5 µm) and intraoocyte reduced glutathione (GSH) content (1.39–1.41 vs 1.00 pixels per oocyte) relative to oocytes matured in N-N-N-N. Somatic cell nuclear transfer (SCNT) embryos derived from the N-N-N-L treatment had significantly (P < 0.05) higher blastocyst formation (53.5%) than embryos derived from Medium-199 (37.4%) and N-N-N-N (41.8%) treatments. Overall, the results demonstrate that maturation of pig oocytes in hypotonic medium with reduced NaCl during the last 11 h of IVM increases the developmental competence of oocytes after PA and SCNT by improving the cytoplasmic microenvironment, including an increased GSH content in IVM oocytes.

Additional keywords: embryonic development, glutathione, perivitelline space.


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