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【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0.05mmol/L(n=39)和0.1mmol/L(n=58)的两种电融合液以及电压分别为1800V/cm(n=58)和2000V/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2000V/cm下降到1800V/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0.05mmol/L的融合液比较,0.1mmol/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。
【Objective】 Explore the project of using human embryo to transfer human embryo. 【Method】 The collection of scrapped prokaryotic fertilized eggs 117. Construction of prokaryotic reconstructed embryos by micromanipulation and electrofusion. Two kinds of electrofusion solutions of 0.05 mmol / L (n = 39) and 0.1 mmol / L (n = 58) were compared with those of electrospray ionization at 1800 V / cm (n = 58) and 2000 V / cm ) On the fusion rate and development of reconstructed zygotes. 【Result】 The success rate of operation was 70.1%, and the fusion success rate was 59.8%. 43 cleavage embryos were obtained, of which 23 (53.5%) had embryos of 6 cells or more. A total of 14 reconstructed embryos developed to 8 cells or more, resulting in 3 blastocysts. There was no significant difference in the rate of cleavage and blastocyst between reconstructed embryo fusion (P> 0.05), but the rate of 6-cell increased significantly (22.2% vs.60.9 %, P <0.05). The fusion rate of 0.1 mmol / L fusion solution increased significantly (39.3% vs.69.0%, P <0.05) compared with the fusion solution with 0.05 mmol / L Ca (superscript 2+) concentration. The cleavage rate, 6-cell rate and blastocyst rate There was no statistical difference (P> 0.05). 【Conclusion】 It is practicable and economical to reconstruct the human nuclear-genomic heterogeneous embryo by using the method of micro-manipulation-electrofusion.