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影響人囊胚形成因素的研究與人胚胎干細(xì)胞建系的初步探討

發(fā)布時(shí)間:2018-06-17 18:31

  本文選題:囊胚 + 內(nèi)細(xì)胞團(tuán); 參考:《天津醫(yī)科大學(xué)》2006年碩士論文


【摘要】:我國大部分生殖中心在行體外受精-胚胎移植(in vitro fertilization,IVF-ET)時(shí)通常應(yīng)用D_3胚胎進(jìn)行移植,然而囊胚移植更符合生理過程,可減少胚胎移植的數(shù)目,降低并發(fā)癥的發(fā)生。為此,我們通過D_3剩余廢棄胚胎的序貫培養(yǎng)得到了一定數(shù)目的囊胚,分析了影響人囊胚形成的因素,并且利用得到的囊胚在人胚胎肺組織成纖維細(xì)胞(human embryonic pulmonary fibroblasts,HEPFs)飼養(yǎng)層上對人胚胎干細(xì)胞(human embryonic stem cells,hESCs)建系進(jìn)行了初步探討。 研究分為三部分:第一部分,影響人囊胚形成因素的研究;第二部分,飼養(yǎng)層的制備與處理;第三部分,人胚胎干細(xì)胞建系的初步探討。 我們將D_3移植后剩余無凍存價(jià)值的胚胎采用序貫培養(yǎng)的方法培養(yǎng)到D_(5-7),觀察囊胚形成的情況及囊胚的質(zhì)量,分析了D_3胚胎的細(xì)胞數(shù)和質(zhì)量分級與囊胚及優(yōu)質(zhì)囊胚形成之間的關(guān)系,給囊胚移植提供理論支持。我們共得到98位患者D_3移植后剩余無凍存價(jià)值的203個胚胎,在患者簽署知情同意書后進(jìn)入本實(shí)驗(yàn)。將胚胎于D_3轉(zhuǎn)移至G2.3中,每天觀察胚胎發(fā)育的情況直至受精D_(5-7)并且對囊胚進(jìn)行分級。共有51個囊胚形成,囊胚形成率為25.12%,優(yōu)質(zhì)囊胚有14個,占所有囊胚的27.45%,全部由Ⅱ~+級以上胚胎形成。32位有囊胚形成的患者有16人獲臨床妊娠,臨床妊娠率為50%,高于66位無囊胚形成患者28.79%的妊娠率(P0.05)。通過分析,我們得出結(jié)論:D_3胚胎細(xì)胞數(shù)與囊胚形成率呈顯著正相關(guān)關(guān)系,Ⅰ-Ⅱ~+級胚胎比Ⅲ-Ⅳ級胚胎囊胚形成率、優(yōu)質(zhì)囊胚形成率高。
[Abstract]:In most reproductive centers in China, D3 embryos are usually used for in vitro fertilization and embryo transfer (IVF-ETT). However, blastocyst transfer is more suitable for physiological process, which can reduce the number of embryo transfer and reduce the incidence of complications. For this reason, we obtained a certain number of blastocysts through sequential culture of the remaining discarded embryos of DST3, and analyzed the factors affecting the formation of human blastocysts. The blastocysts were used to establish human embryonic pulmonary embryonic stem cells hESCs on the feeder layer of human embryonic pulmonary fibroblast cells (HEPFs). The research is divided into three parts: the first part is the study of the factors affecting the formation of human blastocyst; the second part is the preparation and treatment of feeder layer; the third part is the preliminary study on the establishment of human embryonic stem cells. In this paper, the remaining frozen embryo after DSP 3 transplantation was cultured into DSP 5-7 by sequential culture. The formation of blastocyst and the quality of blastocyst were observed, and the relationship between the cell number and quality grading of DSP 3 embryo and the formation of blastocyst and high quality blastocyst were analyzed. To provide theoretical support for blastocyst transplantation. A total of 203 embryos with no cryopreservation value were obtained from 98 patients after DST3 transplantation, and entered the experiment after the patients signed the informed consent form. The embryos were transferred into G2.3, the development of embryos was observed every day until fertilization, and the blastocysts were classified. A total of 51 blastocysts were formed and the blastocyst formation rate was 25.12. There were 14 high-quality blastocysts, accounting for 27.4545 of all blastocysts. The clinical pregnancy rate was 50, which was higher than that in 66 non-blastocyst patients (28.79%). Through analysis, we concluded that there was a significant positive correlation between blastocyst formation rate and blastocyst formation rate. The blastocyst formation rate of 鈪,

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