Alam Md Emtiaj, Iwata Jun, Fujiki Kana, Tsujimoto Yasunori, Kanegi Ryoji, Kawate Noritoshi, Tamada Hiromichi, Inaba Toshio, Sugiura Kikuya, Hatoya Shingo
Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka 598-8531, Japan.
J Vet Med Sci. 2019 Apr 27;81(4):629-635. doi: 10.1292/jvms.18-0335. Epub 2019 Feb 19.
Feline embryo development was examined for 7 days after fertilization using commercially available human media supplemented with 0.3% bovine serum albumin (BSA) or 5% fetal bovine serum (FBS). Cumulus-oocyte complexes were categorized as Grades 1, 2, and 3 according to morphology. Only-One Medium (OM) was used for in vitro culture (IVC) in OM + BSA, OM + FBS, and OM + BSA/FBS, with BSA supplementation for the first 2 days and FBS for the subsequent 5 days. Embryos cultured in Early Culture Medium (1-2 days) and Blastocyst Medium (3-7 days) were defined as EB + BSA and EB + BSA/FBS. The developmental rate until the blastocyst stage of Grade 1 and 2 oocytes cultured in OM + BSA/FBS was higher than for the other groups and was significantly higher than for the OM + BSA and EB + BSA groups (P<0.01). Grade 3 oocytes cultured in OM + BSA/FBS also showed the greatest proportion of blastocyst formation. However, FBS supplementation throughout the IVC period reduced blastocyst number. The percentage of 2 pronuclei after fertilization as well as blastocyst cell number were significantly higher in Grade 1 and 2 than Grade 3 oocytes when cultured in OM + BSA/FBS (P<0.05). These results indicate that commercially available OM supplemented with BSA for the first 2 days of culture and FBS for the subsequent 5 days is suitable for feline embryo development until the blastocyst stage.
在受精后7天内,使用添加了0.3%牛血清白蛋白(BSA)或5%胎牛血清(FBS)的市售人类培养基对猫胚胎发育进行了研究。根据形态将卵丘-卵母细胞复合体分为1级、2级和3级。仅在“唯一培养基”(OM) + BSA、OM + FBS和OM + BSA/FBS中使用OM进行体外培养(IVC),前2天添加BSA,后5天添加FBS。在早期培养基(1 - 2天)和囊胚培养基(3 - 7天)中培养的胚胎分别定义为EB + BSA和EB + BSA/FBS。在OM + BSA/FBS中培养的1级和2级卵母细胞发育至囊胚阶段的发育率高于其他组,且显著高于OM + BSA和EB + BSA组(P<0.01)。在OM + BSA/FBS中培养的3级卵母细胞也显示出最大比例的囊胚形成。然而,在整个IVC期间添加FBS会减少囊胚数量。当在OM + BSA/FBS中培养时,1级和2级卵母细胞受精后2原核的百分比以及囊胚细胞数量显著高于3级卵母细胞(P<0.05)。这些结果表明,在培养的前2天添加BSA,后5天添加FBS的市售OM适合猫胚胎发育至囊胚阶段。