本文選題：玻璃化冷凍 + 玻璃化溶液　； 參考：《昆明醫(yī)學(xué)院》2007年碩士論文

【摘要】： 目的：以普通冷凍麥管為冷凍載體，聯(lián)合冷凍保護(hù)劑乙二醇(ethylene glycol，EG)和二甲亞砜(dimethyl suloxide，DMSO)玻璃化冷凍人早期卵裂期胚胎，比較它們?cè)诓煌瑵舛冉M合下的冷凍效果，以期獲得最佳濃度組合，為臨床提供一種有效的玻璃化溶液。 方法：以人類體外受精發(fā)育至第三天的廢棄卵裂早期胚胎為實(shí)驗(yàn)對(duì)象，以常用的冷凍麥管為載體，冷凍保護(hù)劑EG和DMSO分別取20％和25％兩個(gè)濃度水平，在這兩個(gè)濃度水平EG和DMSO交叉配伍組成不同的玻璃化溶液，分別對(duì)胚胎進(jìn)行玻璃化凍存，并與程序化慢速冷凍法進(jìn)行對(duì)比。胚胎凍存時(shí)間平均為1月，，復(fù)蘇后，通過倒置顯微鏡下形態(tài)學(xué)觀察，比較胚胎的存活率和繼續(xù)發(fā)育能力，另外，復(fù)蘇后存活胚胎經(jīng)羅丹明123(Rhodamine 123)染色，激光掃描共聚焦顯微鏡攝片分析，測(cè)定胚胎線粒體跨膜電位。 結(jié)果：EG和DMSO在20％和25％兩個(gè)濃度水平交叉配伍組成的四種玻璃化溶液E20D20、E20D25、E25D20、E25D25都能有效地凍存人早期卵裂期胚胎，胚胎復(fù)蘇后存活率依次為51.0％、72.7％、62.0％、46.3％，均比程序化慢速冷凍組的存活率39.7％高，其中最高的是20％EG和25％DMSO配伍組，與程序化冷凍組相比差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)，而其余各組與程序化冷凍組相比差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。復(fù)蘇后囊胚形成率依次為11.8％、18.2％、18.0％、9.3％，也均高于程序化冷凍組的囊胚形成率8.6％，其中E20D25組最高，但是各組間的差異均無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。復(fù)蘇后存活胚胎線粒體膜電位的比較，E20D25組膜電位值最高，與未冷凍對(duì)照組相比差異無(wú)統(tǒng)計(jì)學(xué)差異，與其它各組相比，除E25D25組外差異均有統(tǒng)計(jì)學(xué)意義。程序化冷凍組膜電位低于未冷凍對(duì)照組，差異有統(tǒng)計(jì)學(xué)意義(P=0.001＜0.05)。 結(jié)論：1．以EG和DMSO為主體的玻璃化溶液能有效地冷凍人早期卵裂期胚胎。2．以常用的冷凍麥管為載體，20％EG和25％DMSO配伍組成的玻璃化溶液能獲得較好的冷凍效果，可作為臨床應(yīng)用時(shí)一種可供選擇的玻璃化溶液。3玻璃化冷凍技術(shù)比程序化慢速冷凍法能獲得更好的冷凍效果，能夠有效保存人早期卵裂期胚胎。
[Abstract]:Aim: to vitrify human early cleavage embryos with common cryopreservation wheat tube as cryopreservation carrier, combined with ethylene glycol glycoline and dimethyl sulfoxide dimethyl suloxide-DMSO. and compare their freezing effects under different concentration combinations. In order to obtain the best concentration combination, to provide an effective vitrification solution for clinical. Methods: the abandoned early cleavage embryos from the third day of human IVF development were used as experimental objects. The cryopreservation agents EG and DMSO were taken at 20% and 25% concentrations, respectively, using common cryopreserved wheat tube as the carrier, and the concentration of EG and DMSO were 20% and 25%, respectively. At these two concentrations EG and DMSO crosscompatibility composed of different vitrification solution, the embryos were vitrified and frozen, and compared with programmed slow freezing method. The average cryopreservation time was 1 month. After resuscitation, the survival rate and developmental ability of the embryos were compared by morphological observation under inverted microscope. In addition, the surviving embryos were stained with Rhodamine 123(Rhodamine 123 after resuscitation. The transmembrane potential of embryo mitochondria was measured by laser scanning confocal microscope. Results the four vitrified solutions E20D20, E20D25, E20D25, E25D20, E25D20, E25D25 and E25D25 could effectively freeze human early cleavage embryos at 20% and 25% levels, respectively. The survival rate after embryo resuscitation was 51.0% and 72.7%, respectively, which was higher than that of programmed slow freezing group (39.7%). The highest one was 20%EG and 25%DMSO (P < 0.05), but there was no significant difference between the other groups and programmed freezing group (P > 0.05). The blastocyst formation rate after resuscitation was 11.8and 18.2and 18.00.It was also higher than the blastocyst formation rate of programmed freezing group (8.6%). The highest blastocyst formation rate was found in E20D25 group, but there was no significant difference between each group (P > 0.05). Comparison of mitochondrial membrane potential in surviving embryos after resuscitation group E20D25 had the highest membrane potential value, but there was no significant difference compared with other groups except E25D25 group. The membrane potential in programmed freezing group was significantly lower than that in unfrozen control group (P < 0.05). Conclusion 1. Vitrification solution with EG and DMSO as main body can effectively freeze human early cleavage embryos. The glass solution composed of common frozen wheat tube and EG and 25%DMSO can obtain better freezing effect. It can be used as an alternative vitrification technique in clinical application, which can obtain better cryopreservation effect than programmed slow freezing method, and can effectively preserve human early cleavage embryos.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R321-33

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相關(guān)期刊論文 前4條

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