人绒毛膜匀浆及人脐静脉内皮细胞对人未成熟卵母细胞体外成熟的影响

Influences of UPHC culture system and ECHUV co-culture system on the maturation of human immature oocyte in vitro

目的 :研究人早孕绒毛膜匀浆 (UPHC)和人脐静脉内皮细胞 (ECHUV)对人未成熟卵母细胞体外成熟的影响及其作用机理。方法 :应用UPHC培养系统和ECHUV单层细胞培养系统分别培养人未成熟卵母细胞 ,观察卵母细胞体外成熟情况 ;用ELISA方法检测各培养系统中表皮生长因子 (EGF)、粒细胞巨噬细胞集落刺激因子 (GM CSF)和血管表皮生长因子 (VEGF)的浓度。结果 :人未成熟卵母细胞在UPHC和ECHUV培养系统中的成熟率分别为 4 1.0 %和 4 5 .2 % ,明显高于对照组的 14 .3% (P <0 .0 5 ) ;UPHC培养系统和ECHUV培养系统中 ,EGF的浓度分别为 5 2 .6 5± 7.37pg/ml和 5 8.14± 3.0 3pg/ml ,GM CSF的浓度分别为 11.81± 3.76pg/ml和 9.2 0± 2 .5 9pg/ml ,VEGF的浓度分别为 6 1.4 3± 11.38pg/ml和 2 5 6 .5 4± 9.15pg/ml ,均显著高于对照组 (P <0 .0 5 ) ;ECHUV组的VEGF浓度又显著高于UPHC组 (P <0 .0 1) ;<30岁者卵母细胞体外成熟率为 4 2 .6 % ,显著高于 30～ 4 0岁和 >4 0岁的 2 2 .2 %和 13.3% (P <0 .0 5 )。卵 冠 丘复合物体外成熟率为 37.9% ,显著高于裸卵的 2 4 .3% (P <0 .0 5 )。结论 :UPHC和ECHUV促进人未成熟卵母细胞体外成熟 ,其作用与EGF、GM CSF、VEGF有关 ;年龄和卵丘细胞是卵母细胞体外成熟的影响因素。

Objective:To investigate the effect of UPHC and ECHUV culture systems on the maturation of human immature oocyte in vitro,and to explore their active mechanisms and affecting factors.Methods:Human immature oocytes obtained from premenopausal women ovaries were cultured either in UPHC culture system or co cultured with monolayer ECHUV.The maturation rate of the oocytes was evaluated in the two culture systems.The concentrations of epithelial growth factor (EGF),granulocyte macrophage colony stimulating factor (GM CSF) and vascular epithelial growth factor (VEGF) were assayed by enzyme linked immunosorbent assay (ELISA).Results:The maturation rate of human immature oocytes in the two culture systems were 41.0% and 45.2% respectively,which were significantly higher than that of the control group (14.3%,P<0.05).The concentration of EGF in the both culture systems were 52.65±7.37pg/ml and 58.14±3.03 pg/ml,GM CSF were 11.81±3.76pg/ml and 9.20±2.59pg/ml;VEGF were 61.43±11.38pg/ml and 256.54±9.15pg/ml,which were all higher than those of the control group (P<0.05).In addition,the concentration of VEGF in ECHUV system was significantly higher than that of UPHC system (P<0.01).The maturation rate of the oocytes from women whose ages were less than 30 years was 42.6%,which was significantly higher than that of women whose ages were 30～40 years and more than 40 years(22.2% and 13.3%,P<0.05).The maturation rate of ooccyte corona cumulus complex(OCCC) was 37.9%,which was significantly higher than that of the denuded oocytes (24.3%,P<0.05).Conclusions:UPHC culture system and ECHUV co culture system can promote the maturation of human immature oocytes in vitro.Their effects are related to EGF,GM CSF and VEGF.The patient's age and cumulus cells are important factors affecting the maturation rate of human oocyte in vitro.