Miyano T, Hiro-Oka R E, Kano K, Miyake M, Kusunoki H, Kato S
Laboratory of Animal Reproduction, Faculty of Agriculture Kobe University, Nada-ku, Kobe 657, Japan.
Theriogenology. 1994;41(6):1299-305. doi: 10.1016/0093-691x(94)90488-5.
The purpose of this study was to evaluate the ability of hyaluronic acid to improve the development of 1- and 2-cell porcine embryos to the blastocyst stage in a simple medium. In Experiment 1, we confirmed the ability of Whitten's medium supplemented with 15 mg/ml BSA to support the development of porcine embryos to the blastocyst stage under our experimental conditions. Embryos collected from oviducts were cultured at 38.5 degrees C in an atmosphere of 5% CO(2) in humidified air up to 6 d. After 2 d of culture, 82 and 78% of embryos reached the 4-cell stage or beyond in TCM199 supplemented with 10% fetal calf serum (FCS) and in Whitten's medium with BSA, respectively. However, no embryo developed to the morula stage in TCM199 after 6 d of culture. On the other hand, 26 and 15% of embryos developed to the morula and the blastocyst stage in Whitten's medium, respectively. In Experiment 2, we determined whether supplementation of hyaluronic acid in Whitten's medium would improve the development of porcine embryos to the blastocyst stage. After 6 d of culture, development of the embryos to the blastocyst stage was best supported in Whitten's medium with 4 mg/ml BSA and 0.5 mg/ml hyaluronic acid (70%). The proportion of degenerated embryos was lower in the presence than in the absence of hyaluronic acid. These results indicate that the supplementation of Whitten's medium with hyaluronic acid improves the development of 1- and 2-cell porcine embryos to the blastocyst stage.
本研究的目的是评估透明质酸在一种简单培养基中促进猪1细胞和2细胞胚胎发育至囊胚阶段的能力。在实验1中,我们证实了在我们的实验条件下,添加15 mg/ml牛血清白蛋白(BSA)的惠顿氏培养基支持猪胚胎发育至囊胚阶段的能力。从输卵管采集的胚胎在38.5℃、含5%二氧化碳的湿润空气中培养长达6天。培养2天后,添加10%胎牛血清(FCS)的TCM199培养基和含BSA的惠顿氏培养基中分别有82%和78%的胚胎发育至4细胞阶段或更高级阶段。然而,在TCM199培养基中培养6天后,没有胚胎发育至桑椹胚阶段。另一方面,在惠顿氏培养基中分别有26%和15%的胚胎发育至桑椹胚和囊胚阶段。在实验2中,我们确定在惠顿氏培养基中添加透明质酸是否会促进猪胚胎发育至囊胚阶段。培养6天后,在含4 mg/ml BSA和0.5 mg/ml透明质酸的惠顿氏培养基中,胚胎发育至囊胚阶段的情况最佳(70%)。存在透明质酸时退化胚胎的比例低于不存在透明质酸时。这些结果表明,在惠顿氏培养基中添加透明质酸可促进猪1细胞和2细胞胚胎发育至囊胚阶段。