CAJ | 학술논문

目的：以家兔为实验动物模型，探索卵母细胞生发泡(GV)互换的可行性。方法：在注射第一针卵泡刺激素(FSH)72 h后，取大耳白兔卵巢内GV期卵母细胞，用显微操作和电融合的方法，对兔初级卵母细胞进行异卵间GV互换，将重构卵培养于TCM-199加10%FCS中，对重构卵进行成熟培养，最后对成熟重构卵进行卵胞浆内精子注射(ICSI)和胚胎培养。结果：对987枚GV期卵母细胞进行了异卵间GV互换，成功互换272枚(27.6%)，电融合后存活230枚(84.6%)，在进行成熟培养后，230枚卵有178枚(77.4%)发育到MII期(排出第一极体)，将成熟卵进行ICSI后，有126枚(70.8%)受精。结论：对兔初级卵母细胞进行GV互换是可行的，若将此法应用于提高高龄妇女卵母细胞的质量，将有助于解决她们的不育问题。
목적：이가토위실험동물모형，탐색란모세포생발포(GV)호환적가행성。방법：재주사제일침란포자격소(FSH)72 h후，취대이백토란소내GV기란모세포，용현미조작화전융합적방법，대토초급란모세포진행이란간GV호환，장중구란배양우TCM-199가10%FCS중，대중구란진행성숙배양，최후대성숙중구란진행란포장내정자주사(ICSI)화배태배양。결과：대987매GV기란모세포진행료이란간GV호환，성공호환272매(27.6%)，전융합후존활230매(84.6%)，재진행성숙배양후，230매란유178매(77.4%)발육도MII기(배출제일겁체)，장성숙란진행ICSI후，유126매(70.8%)수정。결론：대토초급란모세포진행GV호환시가행적，약장차법응용우제고고령부녀란모세포적질량，장유조우해결저문적불육문제。
Objective: Using rabbit as a model to evaluate the feasibility of the germinal vesicle (GV)transfer. 　　Methods: Collecting the GV stage oocytes from the ovaries of white Japan big ear rabbits at 72 h after the first follicle stimulating hormone injection. The GV-karyoplast was exchanged between the primary oocytes, the pairs of GV karyoplast-cytoplast were fused by electrical pulses, and then the reconstructed oocytes were maturated in TCM-199 with 10% FCS. Finally, the matured reconstructed oocytes were fertilized by intracytoplasmic sperm injection (ICSI) with the same species' sperms. 　　Results:　A total of 987 GV stage oocytes were manipulated. 272 (27.6%) GV-karyoplasts were removed and replaced successfully without lysis. Karyoplast fusion was accomplished in 230 for a final success rate of 84.6% (230/272). After in vitro maturation 178 of 230 (77.4%) reached metaphase II (emitted first polar body). The fertilization rate of matured reconstructed oocytes by ICSI was 70.8% (126/178).Conclusion:　The result indicated that GV transfer was feasible, which could potentially become a new approach in treatment of women infertility due to advanced maternal age.