Chung Jin-Tae, Tosca Lucie, Huang Tian-Hua, Xu Lan, Niwa Koji, Chian Ri-Cheng
Department of Obstetrics and Gynecology, McGill University, Montreal, Canada H3A 1A1.
Reprod Biomed Online. 2007 Aug;15(2):198-207. doi: 10.1016/s1472-6483(10)60709-2.
The exact role of polyvinylpyrrolidone (PVP) in culture medium for oocyte maturation is still largely unknown. Bovine cumulus-oocyte complexes (COC) were cultured in in-vitro maturation (IVM) medium supplemented with 10% fetal bovine serum (FBS), 0.3% PVP (K-90) or 10% serum substitute supplement (SSS) respectively. The rates of oocyte maturation, fertilization and early embryonic development were evaluated. In addition, the status of DNA fragmentation in the oocytes was determined by comet assay, and the ratio of trophectoderm (TE) cells and inner cell mass (ICM) in blastocysts was determined by differential staining. Furthermore, the percentage of apoptotic cells in the blastocysts was examined by TUNEL assay. The results indicated that the effect of PVP in IVM medium was similar to FBS in terms of oocyte maturation and subsequent embryonic development. However, the addition of SSS in IVM medium retarded further embryonic development and resulted in more oocyte DNA fragmentation and a higher ratio of TE cells and ICM in the blastocysts. However, the number of apoptotic cells in blastocysts was similar among the three groups. These results suggest for the first time that the addition of PVP in oocyte maturation medium is not only a suitable substitute for serum but is also beneficial to in-vitro oocyte maturation.
聚乙烯吡咯烷酮(PVP)在卵母细胞成熟培养基中的具体作用仍 largely 未知。牛卵丘 - 卵母细胞复合体(COC)分别在添加 10%胎牛血清(FBS)、0.3% PVP(K - 90)或 10%血清替代补充剂(SSS)的体外成熟(IVM)培养基中培养。评估了卵母细胞成熟、受精和早期胚胎发育的比率。此外,通过彗星试验确定卵母细胞中 DNA 片段化的状态,通过差异染色确定囊胚中滋养外胚层(TE)细胞和内细胞团(ICM)的比例。此外,通过 TUNEL 试验检测囊胚中凋亡细胞的百分比。结果表明,就卵母细胞成熟和随后的胚胎发育而言,IVM 培养基中 PVP 的作用与 FBS 相似。然而,IVM 培养基中添加 SSS 会阻碍进一步的胚胎发育,并导致更多的卵母细胞 DNA 片段化以及囊胚中 TE 细胞与 ICM 的比例更高。然而,三组中囊胚中凋亡细胞的数量相似。这些结果首次表明,在卵母细胞成熟培养基中添加 PVP 不仅是血清的合适替代品,而且有利于体外卵母细胞成熟。