人类卵子体外成熟及其胚胎冷冻复苏研究

In vitro maturation—in vitro fertilization—embryo transfer—frozen-thawed of human oocytes

目的:探讨取卵周期准备、卵泡发育状态、培养液成分对未成熟卵的获取率、体外成熟率、受精率、胚胎质量、胚胎移植后成功率及其胚胎冷冻复苏移植的影响。方法:19例进行卵子体外成熟的不孕不育患者为研究对象。其中1例为自然周期,14例为促性腺激素(Gn)诱导排卵周期,4例常规控制性促排卵(COS)周期;分别以TCM 199或人输卵管液(HTF)为基础培养液,进行卵子体外成熟培养。结果:当取卵时卵泡大小不均一且最大卵泡直径≥12 mm时,获卵率、受精率和胚胎质量下降;TCM 199组优质胚胎的形成率显著高于HTF组(P<0.01)。IVM胚胎经冷冻复苏、胚胎移植能获得与常规IVF周期相似的妊娠成功,出生后代健康、无畸形。结论:在IVM周期取卵时卵泡大小不均一且最大卵泡直径≥12 mm时,将影响胚胎着床率和累积妊娠率。TCM 199优于HTF培养液,能提高未成熟卵体外成熟后受精卵所形成胚胎的发育能力。IVM胚胎经冷冻复苏-胚胎移植能获得与常规IVF周期相似的妊娠成功,出生后代健康、无畸形。

Objective： To investigate the influence of different cycles,ovarian follicle size and IVM culture media on the number of retrieved immature oocytes, maturation rate, fertilization rate ,embryo quality and implantation rate ,pregnancy rate ,delivery rate ,survival and development of frozen-thawed embryos from IVM. Methods： The oocytes were obtained by follicular aspiration from 19 women undergoing oocyte retrieval for in vitro maturation due to the possible risk of ovarian hyperstimulation in IVF-ET program. One patient was in natural cycle, four patients were in ovulation induction cycles with gonadotropine and fourteen patients is controlled ovarian stimulated cycles. All the oocytes retrieved from follicles with 10. 0 - 13. 5 mm in maximumdiameter were allowed to culture in medium M-199 （TCM 199） or HTF supplemented with other substance. Results： When there were nonuniform diameters of follicles and the diameter of largest oocyte exceeded 12 mm,the retrieval rate of oocytes, fertilization rate,and the number of high-quality embryos decreased. The high-quality embryos formation rate was higher for the oocytes cultured in TCM 199 medium than in HTF medium （P〈0. 01）. After being frozen-thawed, the IVM embryos could achieve the same outcome when compared with the conventional IVF treatment. In addition, the offspring were healthy. Conclusions： When the nonuniform diameters of follicles and the diameter of largest oocyte exceeds 12 mm,the retrieval rate of oocytes, fertilization rate,and the number of high-quality embryos decreased. TCM199- based medium is better to improve the developmental potential and implantation rate of embryos derived from in vitro matured oocytes. After being frozen-thawed,the IVM embryos could achieve the are same outcome when compared with the conventional IVF treatment. In addition,the offspring healthy.