Naruse Kenji, Kim Hong Rye, Shin Young Min, Chang Suk Min, Lee Hye Ran, Park Chang Sik, Jin Dong Il
Division of Animal Science and Resources, Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon City 338-708, South Korea.
Theriogenology. 2007 Jan 15;67(2):407-12. doi: 10.1016/j.theriogenology.2006.08.009. Epub 2006 Sep 28.
To investigate the effects of water-soluble vitamin supplementation for IVM/IVC of porcine oocytes and evaluate maturation and developmental capacity in vitro, porcine cumulus oocyte complexes (COCs) was matured in NCSU-23-based medium with water-soluble vitamins for 44 h and then cultured in PZM-3 for 7 days following activation. The COCs were allocated into five treatment groups and matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, 0.4, and 1x). Metaphase II plates of the cumulus-free oocytes were observed following Hoechest 33258 staining. The COCs were allocated into four treatment groups, matured in various concentrations of MEM vitamins (control, 0.05, 0.1, 0.2, and 0.4x) and cultured in PZM-3 following activation. Also, COCS were matured without MEM vitamins and cultured in PZM-3 with various concentrations (control, 0.1, 0.4, 1.0, and 2.0 x) of MEM vitamins. Furthermore, 2 x 2 factorial (IVM/IVC) experiments were performed in IVM medium with or without 0.05 x MEM vitamins and IVC medium with or without 0.4x MEM vitamins to examine the in vitro development of parthenogenetic embryos. Maturation rates of COCs treated with MEM vitamins did not differ significantly among groups. However, compared to the control group, oocytes matured with the addition of 0.05 x MEM vitamins developed to blastocysts at a higher percentage (P<0.05) following activation and culture in PZM-3 without MEM vitamins. Total cell number of blastocysts was significantly higher in the 0.05 x group. Addition of 0.4x MEM vitamins decreased (P<0.05) cleavage and blastocyst developmental rates compared with 0.05 x MEM vitamins-treated group. In contrast, addition of vitamins to PZM-3 medium for in vitro culture of activated porcine oocytes did not affect development. In conclusion, addition of a low concentration of MEM vitamins to IVM medium for porcine oocytes enhanced subsequent development and improved embryo quality.
为研究水溶性维生素补充剂对猪卵母细胞体外成熟/体外培养（IVM/IVC）的影响，并评估其体外成熟和发育能力，将猪卵丘卵母细胞复合体（COCs）在含有水溶性维生素的基于NCSU - 23的培养基中成熟培养44小时，然后在激活后于PZM - 3中培养7天。将COCs分为五个处理组，在不同浓度的MEM维生素（对照、0.05、0.1、0.2、0.4和1倍）中成熟。用Hoechest 33258染色后观察裸卵的中期II板。将COCs分为四个处理组，在不同浓度的MEM维生素（对照、0.05、0.1、0.2和0.4倍）中成熟，并在激活后于PZM - 3中培养。此外，COCs在无MEM维生素的条件下成熟，并在含有不同浓度（对照、0.1、0.4、1.0和2.0倍）MEM维生素的PZM - 3中培养。此外，进行了2×2析因（IVM/IVC）实验，在含或不含0.05倍MEM维生素的IVM培养基以及含或不含0.4倍MEM维生素的IVC培养基中，以研究孤雌胚胎的体外发育。用MEM维生素处理的COCs的成熟率在各组之间无显著差异。然而，与对照组相比，添加0.05倍MEM维生素成熟的卵母细胞在激活并在无MEM维生素的PZM - 3中培养后，发育为囊胚的百分比更高（P<0.05）。0.05倍组的囊胚总细胞数显著更高。与0.05倍MEM维生素处理组相比，添加0.4倍MEM维生素降低了（P<0.05）卵裂率和囊胚发育率。相反，在用于激活猪卵母细胞体外培养的PZM - 3培养基中添加维生素对发育没有影响。总之，在猪卵母细胞的IVM培养基中添加低浓度的MEM维生素可增强后续发育并改善胚胎质量。
