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目的:研究表皮生长因子(epidermalgrowthfactor,EGF)在小鼠早期胚胎体外发育中的作用,以及EGF联合胰岛素样生长因子-Ⅱ(insulin-likegrowthfactor-II,IGF-Ⅱ)对小鼠早期胚胎体外发育是否有协同促进作用。方法:①在mKSOM培养液中添加0ng/ml(对照组)、0.1ng/ml、1ng/ml、10ng/ml、100ng/ml的EGF、10ng/mlIGF-Ⅱ、1ng/mlEGF+1ng/mlIGF-Ⅱ、1ng/mlEGF+10ng/mlIGF-Ⅱ、10ng/mlEGF+1ng/mlIGF-Ⅱ、10ng/mlEGF+10ng/mlIGF-Ⅱ培养小鼠1-细胞胚胎,每组胚胎在培养箱中连续培养120h,每24h观察胚胎发育情况,分别计算2-细胞率、4-细胞率、桑椹胚率、囊胚率和孵化率,并进行囊胚细胞计数。结果:添加1ng/ml、10ng/mlEGF组的囊胚率及孵化率显著增加;EGF+IGF-Ⅱ组较单一添加组、对照组囊胚率及孵化率显著增加(P<0.05),且EGF+IGF-Ⅱ组囊胚细胞计数显著高于其他组(P<0.05),其中10ng/mlEGF+10ng/mlIGF-Ⅱ组的孵化率及囊胚细胞数最高(P<0.05)。结论:EGF浓度在1￣10ng/ml范围内可以提高体外培养鼠胚的囊胚率及孵化率;EGF及IGF-Ⅱ对体外培养鼠胚的发育有协同促进作用;本实验范围内10ng/mlEGF+10ng/mlIGF-Ⅱ为最优组合。
OBJECTIVE: To investigate the role of epidermal growth factor (EGF) in the development of early mouse embryos in vitro and whether EGF combined with insulin-like growth factor-Ⅱ (IGF-Ⅱ) A synergistic role in promoting. Methods: ①In the mKSOM medium, 0ng / ml (control group), 0.1ng / ml, 1ng / ml, 10ng / ml, 100ng / ml EGF, 10ng / mlIGF-Ⅱ, 1ng / mlEGF + 1ng / mlIGF- The embryos were cultured in the incubator continuously for 120 hours, and then cultured in the incubator for 1, 2, 1 ng / ml EGF + 10 ng / ml IGF-II, 10 ng / ml EGF + 1 ng / ml IGF- The development of embryos was observed every 24h. The rates of 2-cell, 4-cell, morula, blastocyst and hatch were calculated and the number of blastocysts was counted. Results: The blastocyst rate and hatching rate of EGF + IGF-Ⅱ group increased significantly (P <0.05) compared with single addition group and control group (P <0.05), and EGF + Blastocyst count in IGF-Ⅱ group was significantly higher than that in other groups (P <0.05). Incubation rate and blastocyst number in 10ng / ml EGF + 10ng / mlIGF-Ⅱ group were the highest (P <0.05). Conclusion: EGF concentration in the range of 1 ~ 10ng / ml can increase the blastocyst rate and hatching rate in vitro; EGF and IGF-Ⅱ have a synergistic effect on the development of mouse embryos cultured in vitro; within the experimental range of 10ng / mlEGF + 10ng / mlIGF-Ⅱ is the best combination.