生长因子对卵母细胞体外成熟及早期胚胎体外发育的影响

EFFECT OF GROWTH FACTORS ON IN VITRO MATURATION OF BOVINE OOCYTES AND THEIR SUBSEQUENT DEVELOPMENT TO BLASTOCYST STAGE

为了探讨生长因子（ＥＧＦ，ＰＤＧＦ）对牛卵母细胞体外成熟与早期胚胎体外发育的影响，本研究进行了两个实验。实验１：在卵母细胞体外成熟（ＩＶＭ）中，将卵母细胞平均随机的放入表皮生长因子浓度为０，２．５，２５，５０μｇ／Ｌ的成熟培养液和对照组中。实验２：在早期胚胎体外培养（ＩＶＣ）过程中，将受精卵平均随机分配入５个不同培养液的处理组：（１）ＴＣＭ－１９９＋１０％阉公牛血清（ＳＳ）；（２）ＴＣＭ－１９９＋ＥＧＦ＋ＰＤＧＦ；（３）ＴＣＭ－１９９＋ＥＧＦ；（４）ＴＣＭ－１９９＋ＰＤＧＦ；（５）ＴＣＭ－１９９＋无生长因子、其中ＥＧＦ：５０μｇ／Ｌ，ＰＤＧＦ：０．１μｇ／Ｌ。２，３，４，５组有０．１％ＰＶＡ和０．３％ＢＳＡ，并与颗粒细胞单层协同培养。卵母细胞的体外成熟（ＩＶＭ）、体外受精（ＩＶＦ）和早期胚胎体外培养（ＩＶＣ）的方法与Ｌｕ等（１９８７）报道的相同。结果表明：浓度为５０μｇ／Ｌ的处理组分裂率与对照组无差异（８９．０％，９２．４％，Ｐ＞０．０５），而浓度为０，２．５，２５μｇ／Ｌ的处理组分裂率均显著低于对照组（７８．６％，８２．３％，８４．３％，Ｐ＜０．０５），在囊胚率和第７ｄ一级胚胎率上各组均无差异。ＥＧＦ在体?

Two experiments were conducted in this study to determine the effect of growth factors (EGF & PDGF ) on in vitro maturation and culture of bovine oocytes to blastocyst stage. In vitro maturation (IVM), fertilization (IVF) and culture (IVC) of bovine oocytes were performed using the same method described by Lu et al (1987).Experiment 1, Bovine oocytes were randomly assigned to the maturation medium which contained 0, 2.5,25,50 μg/L EGF and the control group (TCM-199+10% SS) during IVM.Experiment 2, The cleaved eggs were randomly assigned to five different media: TCM-199 + (1) 10% Steer Serum;(2) EGF+PDGF;(3) EGF;(4) PDGF;(5) no growth factors (EGF: 50 μg/L, PDGF: 0.1μg/L).(2),(3),(4),and (5) treatment groups contained 0. 1% PVA and 0. 3% BSA, all the cleaved eggs were co-cultured with granulosa cell monolayers. The result in Experiment 1 showed that there was no significant difference between control group and the group with 50 μg/L EGF in cleavage rate (92. 4%, 89. 0%,respectively, P>0. 05),while the cleavage rates of the groups with 0, 2. 5, 25 μg/L EGF were significantly lower than that of control group (78. 6 %, 82. 3%, 84. 3 %, respectively,P<0. 05). No differences were found in blastocyst (BL) rate and day 7 grade one embryo rate among all the groups. In Experiment 2, faster development speed and higher rate of blastocysts (BL) were found in group 3 which was significantly different from group 4 (P <0. 05).However, the BL rate of both groups 3 and 4 was still lower than that of control group (P<0. 05).This study suggests that EGF at a level of 50 μg/L can promote both IVM and IVC of bovine oocytes to BL stage, while PDGF has no possitive effect and there is no interaction between EGF and PDGF.