未成熟人卵母细胞的体外成熟培养、受精及胚胎移植研究

发布时间：2017-12-30 20:27

本文关键词：未成熟人卵母细胞的体外成熟培养、受精及胚胎移植研究　出处：《重庆医科大学》2005年硕士论文　论文类型：学位论文

【摘要】：在体外受精与胚胎移植助孕技术(IVF-ET)中需要获得多个优质卵母细胞和胚胎,才能得到较高的妊娠率,因此需要进行常规地控制性超排卵(COH)。在药物诱发超排卵时,由于卵巢中卵泡不同步发育,所采集的卵母细胞约有 15%未达到成熟,过去常因为不能使用而被丢弃。如果能提高未成熟卵的体外成熟和发育能力并加以利用,将增加每个助孕周期的胚胎数量,增加可移植胚胎和冷冻胚胎的数量,增加累积妊娠率,提高助孕成功率。目的: 研究控制性超排卵周期中获得的未成熟卵体外成熟培养(in vitromaturation,IVM)的方法,及未成熟卵的卵丘细胞和培养成熟时间对其成熟、受精及发育能力的影响,建立有效的人卵体外成熟培养系统,探讨 IVM 在人类辅助生殖技术(ART)中的应用及意义。方法: 1、选取接受控制性超排卵周期助孕治疗的不孕夫妇为研究对象。 2、将采集的卵母细胞分为体内成熟卵(MII 期)和未成熟卵两组。未成熟卵又分为带卵丘细胞(卵冠丘复合体,OCCC)和不带卵丘细胞(裸卵,DO)组(均包括 MI 期和 GV 期),分别进行体外成熟培养 24或 48h。重庆医科大学硕士研究生学位论文 4 3、体外成熟卵采用卵胞浆内单精子显微注射(ICSI)授精。体内成熟卵按预定的助孕方案采用 IVF、ICSI 或 IVF+ICSI 受精。 4、取卵第三天优先选择体内成熟卵产生的胚胎进行移植(ET), 其次选择体外成熟卵产生的胚胎移植,剩余胚胎冷冻保存。 5、比较卵丘细胞和体外成熟时间对未成熟卵体外成熟、受精和胚胎发育的影响;比较体内成熟卵和体外成熟卵的受精、卵裂和优质胚胎形成情况;比较 IVM 技术的使用是否提高了助孕结局。结果: 1、70.5%的未成熟卵在 IVM 24h 后成熟,11.2%在 IVM 48h 后成熟, 体外成熟率达 81.7%。 2、未成熟卵中,OCCC 与 DO 组的体外成熟率、受精率、卵裂率和优质胚胎形成率均无明显差异(P0.05);GV 期与 MI 期卵的体外成熟率、受精率、卵裂率和优质胚胎形成率均无明显差异(P0.05), GV 期 DO 卵 24h 成熟率高于 OCCC 卵,差异显著(P0.05);培养 24h 与 48h 成熟的不成熟卵相比较,受精率、卵裂率无明显差异(P0.05),24h 成熟卵优质胚胎形成率更高(28.3%比 12.5%),但差异无统计学意义 (P0.05)。 3、体内成熟卵与体外成熟卵相比较,正常受精率、异常受精率和卵裂率均无明显差异,体内成熟卵优质胚胎形成率更高,差异显著 (P0.005)。 4、比较 2004 年 14 例与 2003 年 17 例体内成熟卵产生可移植胚胎数量2 个的患者,IVM 技术的使用增加了可移植胚胎、冷冻胚胎数量, 减少了周期取消率,提高了助孕成功率。结论: 1、超排卵周期中获得的未成熟卵具有在体外完成成熟和进一步发育成早期胚胎的能力。
[Abstract]:In the technology of in vitro fertilization and embryo transfer (IVF-ET) will need to get a number of high-quality oocytes and embryos, in order to get a higher pregnancy rate, so we need to carry out routine controlled ovarian hyperstimulation (COH). The drug induced ovulation, because ovarian follicles do not develop synchronously. The collected oocytes about 15% cells did not reach maturity, often because you can not use the discarded. If we can improve the in vitro oocyte maturation and development and utilization, will increase the number of embryos per pregnancy cycle, increase the number of embryos and frozen embryos, increase the cumulative pregnancy rate, improve pregnancy rate.
Objective:
In vitro maturation of oocytes of superovulation in the cycle (in vitromaturation IVM) method, and immature oocyte cumulus cells and culture time on the mature mature, fertilization and developmental competence, to establish a culture system in vitro maturation of human oocytes effectively, IVM in human assisted reproductive technology (ART application) and their significance.
Method:
1, the study object is to select the infertile couples who receive the controlled cycle of ovarian hyperstimulation.
2, the collected oocytes were divided into oocytes matured in vivo (MII) and immature oocytes of two groups. The immature eggs are divided into with cumulus cells (OCCC, OCCC) and without (naked oocytes, cumulus cells (DO) group were including MI and GV), respectively, 24 in vitro maturation or 48h.
Master's degree thesis of Medical University Of Chongqing
Four
3, in vitro mature eggs are inseminated by intracytoplasmic sperm injection (ICSI).
Mature eggs are fertilized by IVF, ICSI, or IVF+ICSI according to the predetermined scheme.
4, for third days, oocytes were preferred to select embryos from the mature eggs of the body to be transplanted (ET).
Secondly, the embryo transfer produced by the mature egg in vitro was selected, and the remaining embryos were frozen and preserved.
5, comparison of cumulus cells and in vitro maturation time of oocytes in vitro maturation, fertilization and embryo
The effects of fetal development; comparison of fertilization, cleavage, and quality embryos of mature and in vitro mature eggs
The formation of the fetus; the comparison of the use of IVM technology to improve the outcome of the pregnancy.
Result:
Immature eggs of 1,70.5% mature after IVM 24h, and 11.2% mature after IVM 48h.
In vitro maturation rate is up to 81.7%.
2, in the immature eggs, in vitro maturation rate, fertilization rate, cleavage rate and the rate of OCCC and DO
There was no significant difference in the formation rate of high quality embryos (P0.05), and in vitro maturation of GV and MI eggs
There was no significant difference in rate, fertilization rate, cleavage rate and high quality embryo formation rate (P0.05), GV
The 24h maturation rate of DO eggs was higher than that of OCCC eggs (P0.05), and 24h and 48h were cultured.
There was no significant difference in fertilization rate and cleavage rate between mature and immature eggs (P0.05), 24h
The rate of high quality embryos in mature eggs was higher (28.3% than 12.5%), but the difference was not statistically significant.
(P0.05).
3, the normal fertilization rate, the abnormal fertilization rate and the rate of fertilization were compared with the mature ova in vitro.
There was no obvious difference in cleavage rate. The rate of high quality embryos in mature eggs in vivo was higher, and the difference was significant.
(P0.005).
4, compared with 14 cases in 2004 and 17 cases of mature eggs in 2003 to produce transplantable embryos
In 2 patients, the use of IVM technology increased the number of transplanted embryos, the number of frozen embryos,
It reduces the cycle cancellation rate and improves the success rate of pregnancy.
Conclusion:
1, immature eggs obtained in the superovulation cycle are mature and further developed in vitro.
The ability to develop early embryos.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R321

【共引文献】