表皮生长因子和胎牛血清对猪体细胞克隆胚胎发育的影响

Effect of EGF and FBS on the development of porcine cloned embryos

为了研究胚胎培养液中添加表皮生长因子(EGF)和胎牛血清(FBS)对猪体细胞克隆胚胎发育的影响,试验将卵母细胞成熟培养42h后构建克隆胚,进行电融合激活,再使用6-DMAP(2mmol/L)进行化学辅助激活,然后移入NCSU-23培养基培养,激活后第2天添加0,10,20,30ng/mLEGF,7d后检查囊胚率、囊胚细胞数;激活后第0,2,4,6天添加10%FBS,7d后检查囊胚率、囊胚细胞数。结果表明:2～4细胞阶段添加EGF,添加20ng/mLEGF组的囊胚率显著高于对照组(P<0.05)及其他处理,而囊胚细胞数与对照组无显著差异(P>0.05);在第4天添加FBS组的囊胚率显著高于对照组(P<0.05)和其他处理组,而囊胚细胞数与对照组无显著差异(P>0.05)。结果说明添加20ng/mL的EGF能促进猪体细胞克隆胚胎的发育率,在第4天添加FBS更有利于胚胎的发育。

This study was designed to reveal the effect of EGF and FBS on the development of embryos cloned by porcine somatic cell. After cultured in matured medium, porcine oocytes were ennucleared to construct cloned embryos. Reconstructed embryos were activated, then were treated with 6-DMAP （2 mmol/L for 4 h）. After activation,embryos were cultured in NCSU-23 medium containing 0.4% BSA.EGF（0, 10, 20, 30 ng/mL）and was added to NCSU-23 medium in the 2nd day and the rate of blastocytes and number of cells were counted on 7th day. In another experiment, 10% FBS was added to NCSU-23 medium in the 0, 2^th, 4^th, 6^th day and the rate of blastocytes and number of cells were counted on 7^th day. The results demonstrated that treatments with medium added 20 ng/mL EGF had the higher rate of cloned blastocysts than control and other treatments （P〈0.05） ;but there was no difference in cells of blastocysts; treatments with medium added FBS on 4^th day had the higher rate of cloned blastocysts than that of control and other treatments （P〈0.05） ;but there was no difference in cells of blastocysts. In conclusion,medium added 20ng/ml EGF had an benefit effect on porcine cloned embryo proliferation. Moreover, adding FBS on the 4th day showed better effect on development of cloned embryo.