利用废弃胚胎行人原核移植方法的初步建立

 【目的】 探索利用废弃胚胎进行人原核移植的方案?【方法】 收集废弃多原核受精卵117个?用显微操作-电融合的方法构建原核移植重构胚胎?分别比较钙离子浓度0.05 mmol/L(n=39)和0.1 mmol/L(n=58)的两种电融合液以及电压分别为1 800 V/cm(n=58)和2 000 V/cm(n=20)的两种电场条件对重构合子融合率和发育的影响?【结果】 操作成功率70.1%,融合成功率59.8%,得到43个卵裂胚胎,其中23个(53.5%)6细胞以上胚胎?共14个重构胚发育至8细胞或以上,得到3个囊胚?将电压由2 000 V/cm下降到1 800 V/cm,重构胚胎融合率?卵裂率和囊胚率的差异没有统计学意义(P > 0.05),但6-细胞率明显上升(22.2% vs. 60.9%, P < 0.05)?和钙离子浓度为0.05 mmol/L的融合液比较,0.1 mmol/L的融合液融合率明显上升(39.3% vs. 69.0%,P < 0.05),卵裂率?6-细胞率和囊胚率皆无统计学差异(P > 0.05)?【结论】 通过显微操作-电融合的方法,用废弃胚胎重构人核-质异质的胚胎模型,是可行的?经济的?

Human Pronuclear Transfer with Discarded Triploid Zygotes

[Objective] To establish a micromanipulation-electrofusion procedure for human pronuclear transfer with discarded triploid zygotes. [Materials] A total of 117 oocytes showing 3 pronuclears and 2 polar bodies 16-19 h after in vitro fertilization(IVF), were considered as triploid zygotes and used after informed patient consents. Different fusion media protocols (containing 0.05 mmol/L and 0.1 mmol/L calcium chloride) and electrofusion conditions(1 800 V/cm and 2 000 V/cm voltage) were used to assess their influences on the reconstructed zygote development potential. [Result] The manipulation successful rate was 70.1%. The karyoplast fusion success rate was 59.8%. Forty-three reconstructed zygotes cleaved, among which 23 embryos reached 6-cell stage (53.5% of 2-cell embryos) and 14 developed to 8-cell stage with 3 blastocysts. When the voltage was decreased from 2 000 V/cm to 1 800 V/cm, the fusion rate, cleavage rate, and blastocyst rate between these two groups were not statistically different (P > 0.05), but the 6-cell embryo rate significantly increased (22.2% vs. 60.9%, P=0.049). The fusion rate was significantly increased by increasing the calcium concentration from 0.05 mmol/L to 0.1 mmol/L (39.3% vs. 69.0%, P = 0.014). However, no statistical difference was observed in cleavage rate, the 6-cell rate and the blastocyst rate between the two groups(P > 0.05). [Conclusion] Using discarded triploid zygote to reconstruct diploid zygote with micromanipulation-electrofusion method is a practicable and economical method to get cytoplasmic-nuclear heteroplasmic embryo model.