共培养系统的体细胞类型和状态对猪胚胎早期发育的影响

Effects of Type and State of Co-culture Cells on the In-vitro Development of Porcine Embryos Derived from In-vitro Matured and Fertilized Oocytes

经体外成熟、受精和培养获得猪胚胎 ,采用体细胞共培养研究了体细胞类型和状态对胚胎早期发育的影响。取体外成熟的不同直径卵泡 (>5 m m,2～ 5 mm)卵母细胞的卵丘团 (颗粒细胞团 ) ,培养铺层后与猪受精卵共培养 ,组间受精卵的卵裂率和发育能力无显著差异 ;根据卵巢的状况对猪输卵管上皮细胞 (POECs)的状态进行分组 ,受精卵和卵巢表面布满卵泡的 POECs共培养 ,卵裂率显著低于卵巢表面有黄体和 /或红体的 POECs共培养组 (P<0 .0 5 ) ,虽然各组间 3～ 4-细胞的发育率无显著差异 ,但卵巢表面有红体和卵泡的 POECs共培养组的 >4-细胞的发育率显著高于卵巢表面布满卵泡的 POECs共培养组 (P<0 .0 5 ) ;受精卵在共培养系统和非共培养系统中的卵裂率无显著差异 ,受精卵非共培养系统中 3～ 4-细胞的发育能力显著低于颗粒细胞共培养组 (P<0 .0 5 ) ,极显著低于 POECs共培养组 (P<0 .0 1) ,无能力突破 4-细胞继续发育 ,与颗粒细胞单层、POECs单层共培养的受精卵 >4-细胞的发育率分别为 2 4.0 %、5 3.8% ,差异显著 (P<0 .0 5 )。结果表明 ,共培养系统对胚胎体外发育的作用 ,一方面与体细胞类型有关 。

The aim of the present study was to investigate the effects of type and state of co-culture cells on the in-vitro development of porcine embryos derived from in-vitro matured and fertilized oocytes.When zygotes were co-cultured with granulosa cells(GCs) derived from IVM oocytes aspirated from different size follicles(2-5 mm and >5 mm),the cleavage rates and the proportions of 3-4 cell embryos and >4-cell embryos were not significantly different between the groups.Based on the status of ovaries,the state of porcine oviductal epithelial cells(POECs) was grouped.When zygotes were co-cultured with POECs,of which ovaries enriched with follicles in the surface,the cleavage rate was significantly lower than those co-cultured with POECs,of which ovaries were occupied with corpus haemorrhagicum and occupied with corpus luteum and follicles(P<0.05).Although the proportion of 3-4-cell embryos was not significantly different between the groups,the proportion of >4-cell embryos co-cultured with POECs,of which ovaries were occupied with corpus haemorrhagicum and follicles,was significantly higher than that co-cultured with POECs,of which ovaries enriched with follicles(P<0.05).The cleavage rate of oocytes used for IVF was not significantly different among co-culture systems with GCs,with POECs and non-coculture system,but the proportion of 3-4-cell embryos in the non-coculture system was significantly lower than those in the co-culture systems(P<0.05 and P<0.01).Embryos in the non-coculture system showed a complete failure to develop beyond the 4-cell stage,but the proportions of >4-cell embryos co-cultured with GCs and POECs were 24.0% and 53.8%,respectively,and the difference between them was significant(P<0.05).In conclusion,the potential competence to support embryos development is part of difference in both the type of somatic cells as well as the state of POECs cells during co-cultures system.