Abstract

Despite the application of numerous supplements to improve in vitro culture (IVC) conditions of mammalian cells, studies regarding the effect of fetal bovine serum (FBS) on mammalian early embryogenesis, particularly in relation to redox homeostasis, are lacking. Herein, we demonstrated that early development of in vitro-produced (IVP) porcine embryos highly depends on the combination of FBS supplementation timing and embryonic reactive oxygen species (ROS) requirements. Interestingly, FBS significantly reduced intracellular ROS levels in parthenogenetically activated (PA) embryos regardless of the developmental stage. However, the beneficial effect of FBS on early embryogenesis was found only during the late phase (IVC 4–6 days) treatment group. In particular, developmental competence parameters, such as blastocyst formation rate, cellular survival, total cell number and trophectoderm proportion, were markedly increased by FBS supplementation during the late IVC phase. In addition, treatment with FBS elevated antioxidant transcript levels during the late IVC phase. In contrast, supplementation with FBS during the entire period (1–6 days) or during the early IVC phase (1–2 days) greatly impaired the developmental parameters. Consistent with the results from PA embryos, the developmental competence of in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) embryos were markedly improved by treatment with FBS during the late IVC phase. Moreover, the embryonic stage-specific effects of FBS were reversed by the addition of an oxidant and were mimicked by treatment with an antioxidant. These findings may increase our understanding of redox-dependent early embryogenesis and contribute to the large-scale production of high-quality IVP embryos.

Highlights

  • Pigs are considered useful experimental animals for biomedical research, such as xenotransplantation and new-drug discovery, due to their high similarity to humans in anatomical and physiological features [1]

  • To study the effect of fetal bovine serum (FBS) on the early development of porcine embryos, parthenogenetically activated (PA) embryos were cultured in in vitro culture (IVC) medium supplemented with 10% FBS for various culture periods, and the blastocyst development rate and total cell number were assessed after 6 days of IVC

  • Consistent with this, the embryonic reactive oxygen species (ROS) level was successfully decreased by adding 1 mM GSH, which led to significant decreases in cleavage rate, blastocyst development rate, TE cell number, and cellular survival compared with controls (S3 Fig, S5 and S6 Tables)

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Summary

Introduction

Pigs are considered useful experimental animals for biomedical research, such as xenotransplantation and new-drug discovery, due to their high similarity to humans in anatomical and physiological features [1]. For this reason, in vitro-produced (IVP) porcine embryos have been used extensively to generate transgenic animals used as research materials, such as bio-organs and disease models [2,3,4]. Unlike in vivo embryos, poor developmental competence is a major obstacle in the mass production of high-quality porcine IVP embryos To address this issue, many researchers have attempted to improve the quality of porcine IVP embryos by adding numerous supplements to in vitro culture (IVC) medium to reduce the developmental damages caused by in vitro conditions [5,6,7]. Studies regarding the effects of serum supplementation on embryonic stage and the underlying mechanism(s) are lacking

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