Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
Department of Veterinary Physiology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
J Cell Physiol. 2018 Sep;233(9):6984-6995. doi: 10.1002/jcp.26489. Epub 2018 Mar 25.
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.
在不同物种的体外成熟（IVM）中，广泛使用补充剂，如猪卵泡液（pFF）、胎牛血清和人血清白蛋白，但这些补充剂含有未定义的成分，这导致了标准化的技术困难，并影响了 IVM 的效率。无血清替代物（KSR）是一种合成的蛋白质来源，不含任何未定义的生长因子或促进分化的因子。因此，使用 KSR 作为一种确定的成分来避免 IVM 系统中未知分子的影响是可行的。在这项研究中，与未添加组相比，经孤雌激活（PA）、体细胞核移植（SCNT）和体外受精（IVF）后的卵母细胞成熟率和囊胚形成率在 5%KSR 补充组显著提高，并且与 10%pFF 补充组更为相似。此外，GDF9、BMP15、ROS、GSH、BODIPY-LD、BODIPY-FA 和 BODIPY-ATP 染色的强度在 5%KSR 和 10%pFF 之间显示出相似的值，与对照组有显著差异。与两种补充剂相关的大多数与脂质代谢相关的基因表达表现出相似的模式。总之，5%KSR 上调了脂质代谢，从而为 PA、SCNT 和 IVF 后的卵母细胞质量和随后的胚胎发育提供了必需的能量来源。这些迹象支持了这样一种观点，即 KSR 作为卵母细胞 IVM 的定义血清补充剂可能在其他物种中普遍使用。
