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J Cell Physiol. 2018 Sep;233(9):6984-6995. doi: 10.1002/jcp.26489. Epub 2018 Mar 25.

A potential role of knockout serum replacement as a porcine follicular fluid substitute for in vitro maturation: Lipid metabolism approach.

Author information

1
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
2
Department of Veterinary Physiology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
3
Designed Animal Resource Center and Biotransplant Research Institute, Seoul National University Green-Bio Research Complex, Gangwon-do, Korea.
4
Division of Nephrology, Seoul National University College of Medicine, Seoul, Korea.
5
Transplantation Center, Seoul National University Hospital, Seoul, Korea.

Abstract

The use of supplements, such as porcine follicular fluid (pFF), fetal bovine serum and human serum albumin are widely used during in vitro maturation (IVM) in different species but these supplements contain undefined components that cause technical difficulties in standardization and influence the efficiency of IVM. Knockout serum replacement (KSR) is a synthetic protein source, without any undefined growth factors or differentiation-promoting factors. Therefore, it is feasible to use KSR as a defined component for avoiding effects of unknown molecules in an IVM system. In this study, the rates of oocyte maturation and blastocyst formation after parthenogenetic activation (PA), somatic cell nuclear transfer (SCNT) and in vitro fertilization (IVF) were significantly higher in the 5% KSR supplemented group than in the unsupplemented control group and more similar to those of the 10% pFF supplemented group. Moreover, the intensity of GDF9, BMP15, ROS, GSH, BODIPY-LD, BODIPY-FA, and BODIPY-ATP staining showed similar values between 5% KSR and 10% pFF, which have significant difference with control group. Most of the gene expression related to lipid metabolism with both supplements exhibited similar patterns. In conclusion, 5% KSR upregulated lipid metabolism and thereby provides an essential energy source to sustain and improve oocyte quality and subsequent embryo development after PA, SCNT, and IVF. These indications support the idea that KSR used as a defined serum supplement for oocyte IVM might be universally used in other species.

KEYWORDS:

embryo development; knockout serum replacement; lipid metabolism; oocyte maturation; porcine follicular fluid

PMID:
29345310
DOI:
10.1002/jcp.26489

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