【摘要】： 目的:觀察不同培養(yǎng)條件下小鼠胚胎培養(yǎng)液中瘦素的分泌情況,分析培養(yǎng)條件對(duì)胚胎的影響。應(yīng)用小鼠移植模型觀察胚胎自分泌瘦素水平及形態(tài)學(xué)評(píng)分與著床及妊娠的關(guān)系,探討瘦素水平對(duì)胚胎質(zhì)量及其種植潛能的預(yù)測(cè)價(jià)值。 方法:將小鼠2-細(xì)胞期胚胎在三種不同培養(yǎng)條件下分別培養(yǎng)至4-細(xì)胞期、8-細(xì)胞期、桑葚胚及囊胚期:(1)單純序貫培養(yǎng)組;(2)共培養(yǎng)組-序貫培養(yǎng)組;(3)體內(nèi)發(fā)育組。將發(fā)育的同期胚胎(2-細(xì)胞期、4-細(xì)胞期、8-細(xì)胞期、桑葚胚及囊胚期)分別在50ul微滴中培養(yǎng)6小時(shí),移出胚胎后收集微滴。應(yīng)用ELISA法測(cè)定瘦素分泌情況。囊胚供移植用,11日后觀察其受孕情況,比較瘦素水平與形態(tài)學(xué)評(píng)分對(duì)胚胎著床能力的預(yù)測(cè)效果。 結(jié)果:1、共培養(yǎng)-序貫培養(yǎng)組8-細(xì)胞期之后胚胎的發(fā)育速度、囊胚形成率均高于單純序貫培養(yǎng)組(P0.05)。2、三組桑葚胚之前的培養(yǎng)微滴中均未檢測(cè)到瘦素的分泌,但在桑葚胚及囊胚5胚/滴的培養(yǎng)微滴中可檢測(cè)到。其中體內(nèi)發(fā)育組明顯高于單純序貫培養(yǎng)組和共培養(yǎng)-序貫培養(yǎng)組(P0.05),后兩組之間無(wú)顯著性差異(P0.05)。3、形態(tài)學(xué)上評(píng)分相同的胚胎瘦素高分泌組著床率明顯高于低分泌組(P0.01);瘦素高分泌的3BB組囊胚著床率高于低分泌的3AA組囊胚(P0.05)。 結(jié)論:1.共培養(yǎng)-序貫培養(yǎng)組較單純序貫培養(yǎng)組有更高的囊胚形成率,共培養(yǎng)-序貫培養(yǎng)對(duì)胚胎發(fā)育的促進(jìn)作用在8-細(xì)胞期胚以后更明顯。2.胚胎在桑葚胚以后分泌瘦素增多,囊胚期胚胎分泌量明顯增加,且分泌量的高低與胚胎活力有關(guān),可反映胚胎的成熟度和質(zhì)量。3、胚胎自分泌瘦素對(duì)胚胎種植潛能有預(yù)測(cè)價(jià)值,特別是在形態(tài)學(xué)上評(píng)分相同胚胎的選擇上。瘦素對(duì)胚胎種植潛能的預(yù)測(cè)價(jià)值可能優(yōu)于形態(tài)學(xué)評(píng)分,結(jié)合瘦素檢測(cè)和形態(tài)學(xué)評(píng)分來(lái)綜合判斷胚胎著床能力將更有價(jià)值。
[Abstract]:Aim: to observe the secretion of leptin in mouse embryo culture medium under different culture conditions and analyze the effect of culture conditions on embryo. In order to explore the predictive value of leptin level on embryo quality and implanting potential, the relationship between leptin level and implantation and pregnancy was observed in mouse model of transplantation to observe the relationship between leptin level and embryo autosecretory level and morphological score. Methods: mouse 2-cell embryos were cultured to 4-cell stage, 8-cell stage, morula and blastocyst stage under three different culture conditions: (1) sequential culture group; (2) co-culture group-sequential culture group; (3) in vivo development group. The developmental embryos (2-cell stage, 4-cell stage, 8-cell stage, morula stage and blastocyst stage) were cultured in 50uls microdroplets for 6 hours, and the microdroplets were collected after they were removed from the embryos. Leptin secretion was measured by ELISA. The blastocyst was used for transplantation and the pregnancy was observed 11 days later. The effects of leptin level and morphological score on the ability of embryo implantation were compared. Results: 1. The rate of embryo development and blastocyst formation in the co-culture-sequential culture group after 8-cell stage was higher than that in the simple sequential culture group (P0.05). 2. No leptin secretion was detected in the pre-culture microdroplets of the three groups of morulae, and the rate of blastocyst formation was higher than that in the control group (P0.05). However, it can be detected in the culture microdroplets of morula and blastocyst 5 embryos / drop. The in vivo development group was significantly higher than that in sequential culture group and co-culture-sequential culture group (P0.05), but there was no significant difference between the latter two groups (P0.05). The implantation rate in the high-secretion group was significantly higher than that in the low-secretion group (P0.01). The blastocyst implantation rate in 3BB group with high leptin secretion was higher than that in low-secretion 3AA group (P0.05). Conclusions: 1. The rate of blastocyst formation in the co-culture-sequential culture group was higher than that in the simple sequential culture group, and the effect of co-culture-sequential culture on the development of embryos was more obvious after the 8-cell embryo stage. 2. The secretion of leptin by embryos after morula is increased, and the secretion of blastocyst stage embryos is significantly increased, and the level of secretion is related to the activity of embryos, which can reflect the maturity and quality of embryos. Embryonic autocrine leptin has predictive value for embryo implantation potential, especially in morphological evaluation of the same embryo selection. The predictive value of leptin in embryo implantation potential may be superior to morphological score. It will be more valuable to judge embryo implantation ability by combining leptin detection and morphological score.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R321

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相關(guān)期刊論文 前1條

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