生殖医学杂志
生殖醫學雜誌
생식의학잡지
JOURNAL OF REPRODUCTIVE MEDICINE
2001年
3期
141-147
,共7页
王敏康%陈大元%刘冀珑%李光鹏%廉莉%宋祥芬%孙青原
王敏康%陳大元%劉冀瓏%李光鵬%廉莉%宋祥芬%孫青原
왕민강%진대원%류기롱%리광붕%렴리%송상분%손청원
小鼠%卵母细胞%MII期核(纺锤体)移植%电融合%体外受精
小鼠%卵母細胞%MII期覈(紡錘體)移植%電融閤%體外受精
소서%란모세포%MII기핵(방추체)이식%전융합%체외수정
目的:为研究和了解细胞核与细胞质的相互作用。方法:采用3%蔗糖预处理显示及去核法进行昆明白小鼠与C57BL/6小鼠(黑色)之间卵母细胞MII期核(纺锤体)的移植、电融合、体外受精-胚胎移植(IVF-ET)。结果:经3%蔗糖预处理卵母细胞的去核率达100%。完成MII期核移植的重构卵母细胞约17.6%完成电融合。IVF的受精率为82%。受精卵2-细胞胚胎的发育率为80%~90%,8个1~4细胞胚胎移入两只自然怀孕1 d的寄母小鼠(Km×Km)输卵管,其中1只孕鼠生下3只小鼠,其中2只黑色小鼠为MII期核(纺锤体)移植生产的小鼠。移植产仔率为25%。结论:实验结果证明了3%蔗糖预处理显示MII期核(纺锤体)及去核效果可达100%;电融合的重构卵母细胞可完成正常的IVF。为进行不同年龄和卵龄间的MII期核移植提供了一个模型。
目的:為研究和瞭解細胞覈與細胞質的相互作用。方法:採用3%蔗糖預處理顯示及去覈法進行昆明白小鼠與C57BL/6小鼠(黑色)之間卵母細胞MII期覈(紡錘體)的移植、電融閤、體外受精-胚胎移植(IVF-ET)。結果:經3%蔗糖預處理卵母細胞的去覈率達100%。完成MII期覈移植的重構卵母細胞約17.6%完成電融閤。IVF的受精率為82%。受精卵2-細胞胚胎的髮育率為80%~90%,8箇1~4細胞胚胎移入兩隻自然懷孕1 d的寄母小鼠(Km×Km)輸卵管,其中1隻孕鼠生下3隻小鼠,其中2隻黑色小鼠為MII期覈(紡錘體)移植生產的小鼠。移植產仔率為25%。結論:實驗結果證明瞭3%蔗糖預處理顯示MII期覈(紡錘體)及去覈效果可達100%;電融閤的重構卵母細胞可完成正常的IVF。為進行不同年齡和卵齡間的MII期覈移植提供瞭一箇模型。
목적:위연구화료해세포핵여세포질적상호작용。방법:채용3%자당예처리현시급거핵법진행곤명백소서여C57BL/6소서(흑색)지간란모세포MII기핵(방추체)적이식、전융합、체외수정-배태이식(IVF-ET)。결과:경3%자당예처리란모세포적거핵솔체100%。완성MII기핵이식적중구란모세포약17.6%완성전융합。IVF적수정솔위82%。수정란2-세포배태적발육솔위80%~90%,8개1~4세포배태이입량지자연부잉1 d적기모소서(Km×Km)수란관,기중1지잉서생하3지소서,기중2지흑색소서위MII기핵(방추체)이식생산적소서。이식산자솔위25%。결론:실험결과증명료3%자당예처리현시MII기핵(방추체)급거핵효과가체100%;전융합적중구란모세포가완성정상적IVF。위진행불동년령화란령간적MII기핵이식제공료일개모형。
Objective: To study the interaction between nucleus and cytoplasm.
Methods: Operation medium containing 3% sucrose was used to pretreatment murine oocytes and meiotic apparatus transfer(MAT) was carried out from C57BL/6 mouse (black) to enucleated Kunming murine oocytes and followed by electrofusion, in vitro fertilization (IVF) and embryo transfer.
Results: The results showed that the enucleation rate by treating oocytes with 3% sucrose was 100%, 17.6% of reconstructed karyoplasm-recipient cytoplasm pairs fused sucessfully. When the fused oocytes were exposed to spermatozoa from C57BL/6 mice, 9 out of 11 (82%) were fertilized. Among them, eight embryos at one-to four-cell stages were transferred into the oviducts of two synchronous pregnant Kunming strain fosters and one delivered two normal C57BL/6 offsprings.
Conclusion: This study indicated that MII meiotic apparatus or spindle sustains normal structure and function after micromanipulation and electrofusion. MAT provides a model for further research on the application of this technique to assistant human reproduction.