本文關(guān)鍵詞：小鼠囊胚玻璃化冷凍研究及超微結(jié)構(gòu)觀察　出處：《昆明醫(yī)學(xué)院》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 囊胚 冷凍保存 玻璃化 乙二醇 二甲亞砜 超微結(jié)構(gòu)

【摘要】：目的 研究由兩種冷凍保護劑乙二醇(Ethylene Glycol，EG)和二甲亞砜(dimethylsuphoxide，DMSO)單獨或聯(lián)合配伍的改良玻璃化溶液對小鼠囊胚超快速冷凍保存的效果；比較聚蔗糖(Ficoll 70)的添加對超快速冷凍的作用；初步確定囊胚冷凍保存最佳玻璃化溶液及玻璃化冷凍的最佳囊胚發(fā)育階段；通過透射電鏡觀察找出囊胚冷凍損傷的超微結(jié)構(gòu)改變。 方法 1、對6-8W齡昆明小白鼠促排卵，處死小鼠后獲得小鼠初級囊胚和擴張囊胚。2、分別采用六組玻璃化溶液(ES35、EDS35、ESF10、ESF20、EDSF10和EDSF20)對小鼠初級囊胚和擴張囊胚進行玻璃化冷凍。3、透射電鏡下觀察新鮮擴張囊胚和各冷凍組凍融擴張囊胚的超微結(jié)構(gòu)。 結(jié)果 1、對于初級囊胚的存活率和孵出率均是EDSF10組最高，與新鮮對照組無顯著性差異；ES35組最低與EDSF10組和新鮮對照組之間均存在顯著性差異；2、對于擴張囊胚存活率各實驗組間均無顯著性差異，與新鮮對照組均有顯著性差異；對于擴張囊胚的孵出率ES35組最低，EDSF10組最高，與新鮮對照組均存在顯著差異。3、EG(?)DMSO對于囊胚的存活率和孵出率均有統(tǒng)計學(xué)意義。4、Ficoll70對于初級囊胚的存活率有統(tǒng)計學(xué)意義，濃度為10％或20％時的存活率均顯著高于濃度為0％時，濃度為10％時存活率高于20％時，，但無統(tǒng)計學(xué)意義。5、除ES35組外各冷凍組，初級囊胚存活率顯著高于擴張囊胚；EDS35組、ESF20組和EDSF20組的初級囊胚孵出率均顯著高于擴張囊胚孵出率。6、透射電鏡觀察顯示擴張囊胚冷凍損傷主要是細(xì)胞膜性結(jié)構(gòu)和細(xì)胞骨架結(jié)構(gòu)的損傷。 結(jié)論 1、DMSO的添加對于改善玻璃化溶液具有重要作用，尤其是有助于初級囊
[Abstract]:Objective to study two kinds of cryopreservation agents, ethylene glycol Glycoline (EGG) and dimethyl sulfoxide dimethylsuphoxide (DMSO). Effects of modified vitrification solution alone or in combination with DMSO on ultrafast cryopreservation of mouse blastocysts; The effects of polysucrose Ficoll 70 on ultra-fast freezing were compared. The optimum vitrification solution for cryopreservation of blastocyst and the optimum stage of blastocyst development for vitrification were preliminarily determined. The ultrastructural changes of blastocyst cryopreservation injury were observed by transmission electron microscope (TEM). Methods 1. Primary blastocyst and expanded blastocyst were obtained from 6-8 W old Kunming mice. Six groups of vitrified solution (ES35) EDS35 were used respectively to obtain mouse primary blastocyst and dilated blastocyst. ESF10 ESF20 EDSF10 and EDSF20) vitrified mouse primary blastocyst and expanded blastocyst. The ultrastructure of fresh expanded blastocysts and freeze thawed blastocysts were observed under transmission electron microscope. Results 1. The survival rate and hatching rate of primary blastocysts in EDSF10 group were the highest, and there was no significant difference between the two groups. There was significant difference between ES35 group and EDSF10 group and fresh control group. 2. There was no significant difference in blastocyst survival rate between each experimental group and fresh control group. The hatching rate of expanded blastocysts in ES35 group was the highest, which was significantly different from that in fresh control group. DMSO has statistical significance for survival rate and hatching rate of blastocyst. 4 Ficoll70 has statistical significance for survival rate of primary blastocyst. The survival rate at the concentration of 10% or 20 was significantly higher than that at the concentration of 0, and the survival rate at the concentration of 10 was higher than that at the time of 20, but there was no statistical significance. 5. The survival rate of the freezing group was higher than that of the control group except the ES35 group. The survival rate of primary blastocyst was significantly higher than that of expanded blastocyst. The hatching rate of primary blastocysts in EDS35 group and EDSF20 group was significantly higher than that in expanded blastocyst group. Transmission electron microscopy (TEM) showed that the freeze injury of expanded blastocyst was mainly caused by membrane structure and cytoskeletal structure. Conclusion 1 the addition of DMSO plays an important role in improving vitrification solution, especially in primary sac.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R321-33

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