卵巢组织玻璃化冷冻研究进展     

吴志南  丁家桐 《动物医学进展》2004,25(6):14-15

卵巢组织玻璃化冷冻可替代直接冷冻卵母细胞或胚胎。玻璃化冷冻卵巢组织在辅助生殖上具有优越性。它无需控制供体的生殖周期 ,也无需取出卵泡。同样这一技术可用于保存濒危动物或受意外伤害的人或动物的卵母细胞 ;可为性成熟前失去生殖能力的动物或人提供生殖保险以及增加卵母细胞的来源 ;可用于建立生殖细胞 (卵母细胞 )的冷冻库。而传统的冷冻技术存在很多弊端。文章综述了玻璃化冷冻卵巢组织的研究背景和现状 ,并指出了其广阔的应用前景  相似文献   

细胞松弛素B对牛GV期卵母细胞玻璃化冷冻效果的影响     

郭娟  王志琴  赵云程  林嘉鹏  黄俊成 《中国畜牧兽医》2009,36(9):101-103

本试验通过玻璃化冷冻前不同浓度细胞松弛素B(cytochalasin-B,CB)预处理来分析CB对牛GV期卵母细胞固体表面玻璃化(SSV)冷冻后发育潜力的影响。试验结果表明,玻璃化冷冻后,CB处理组和未处理组之间卵母细胞成熟率差异不显著(P>0.05),但均显著低于体外培养组。说明试验中所用CB浓度 (2.5、7.5、15、20和30 μg/mL)对牛GV期卵母细胞玻璃化冷冻效果的改善作用不明显。  相似文献   

库车小白杏的组织培养及玻璃化法超低温保存试验初探     

张娟  王新建  闫福军 《新疆农业科学》2011,48(12):2255-2260

[目的]采用玻璃化法对杏资源茎尖进行超低温保存.超低温保存是目前植物种质资源长期稳定保存的最好方法,保存后的材料遗传上较为稳定.[方法]以库车小白杏为试材,对茎尖(外植体)组织培养的培养基及激素浓度做了初步选择.[结果]选择MS +0.5NAA +0.2 6 -BA激素配方可使杏的茎尖最易形成愈伤材料,而选择MS +0.1 NAA +0.1 6- BA +0.1GA激素配方可使由茎尖获得的愈伤材料最易分化出根.[结论]用玻璃化法对组织培养获得的愈伤材料结合不同的低温保存时间、脱水处理时间与化冻方法进行比较试验可知,预冻温度- 30℃、玻璃化液保护剂脱水60 min、40℃水浴5 min解冻,可使超低温保存的材料复活率最高.  相似文献   

枣子叶再生植株研究   总被引：1，自引：0，他引：1  

郭敬丽  王玖瑞  刘孟军  代丽 《河北农业大学学报》2009,32(4)

以枣树‘雄性不育3号’未成熟胚的子叶为材料,建立了枣子叶再生植株体系。子叶愈伤诱导培养基为MS+蔗糖30 g/L+琼脂3.3 g/L,愈伤诱导率达到40%。从愈伤组织诱导不定芽的培养基为MS+蔗糖30 g/L+琼脂3.3 g/L+TDZ 1.5 mg/L,不定芽发生率为88.9%,不定芽数量与愈伤数量之比为3.5。生根培养基为MS+蔗糖30 g/L+琼脂3.0 g/L+IBA 1.0 mg/L,生根率达到33.3%。此外,培养温度显著影响玻璃化苗的生长。在24℃条件下培养30 d,仅有9.7%的玻璃化苗向正常植株转化,而在20℃培养条件下的玻璃化率仅为6.7%。附加30~80 g/L蔗糖的不同培养基上,玻璃化苗向正常苗转化情况无显著差异。  相似文献   

不同冷冻方法对牛体外胚胎ATP含量与ROS水平的影响   总被引：1，自引：1，他引：0  

赵学明  杜卫华  王栋  郝海生  朱化彬 《中国农业科学》2012,45(1):170-177

【目的】探讨冷冻保存对牛体外生产胚胎能量代谢、有氧代谢造成的影响。【方法】采用常规法或玻璃化法（open pulled straw,OPS法）冷冻保存牛体外受精（in vitro fertilization,IVF）囊胚和体细胞核移植（somatic cell nuclear transfer,SCNT）囊胚,利用ATP Bioluminescence Assay Kit HS II 和 GENMED ROS Assay Kit检测了冷冻-解冻后囊胚ATP含量和ROS水平。【结果】（1）IVF和SCNT囊胚OPS法冷冻-解冻后存活率（（93.25±5.17）%和（77.56±3.52）%）均显著高于常规冷冻法（（81.25±4.98）%和（49.41±2.24）%）（P＜0.05）;（2）IVF囊胚和SCNT囊胚OPS法冷冻-解冻后ATP含量（（0.63±0.05）和（0.33±0.02）pmol）均显著高于常规冷冻法（（0.45±0.03）和（0.22±0.01）pmol）（P＜0.05）;但是均显著低于相应的IVF或SCNT新鲜囊胚（（0.76±0.04）和（0.40±0.02）pmol）（P＜0.05）;（3）玻璃化冷冻IVF囊胚、SCNT囊胚ROS水平（（74.34±4.24）和（43.21±3.35）cps）高于新鲜IVF囊胚（（48.52±2.65）cps）、SCNT囊胚（（27.36±2.23）cps）（P＜0.05）,常规冷冻组则与此相反（（35.61±4.32）和（16.56±2.52）cps）（P＜0.05）。【结论】玻璃化冷冻较适用于牛IVF、SCNT囊胚冷冻保存;玻璃化冷冻和常规冷冻均显著影响牛IVF囊胚、SCNT囊胚中ATP含量和ROS水平。  相似文献   

高良姜种子超低温保存研究   总被引：2，自引：1，他引：1  

曾琳  何明军  陈葵  郑希龙 《中国农学通报》2014,30(28):164-168

为解决高良姜种子长期保存的问题,以高良姜的成熟种子为材料,研究含水量和玻璃化处理对其活力的影响,以及快速冷冻法和玻璃化冷冻法对种子超低温保存的影响。结果表明：（1）种子含水量在10.77%~27.25%时,其活力仍保持在60%以上,当含水量由10.77%降至7.86%时,其活力降至40%以下;（2）玻璃化处理后的种子活力低于未玻璃化处理的种子活力;（3）经液氮超低温保存后,含水量为21.10%~27.25%的种子活力降至50%以下甚至为0%;含水量为10.77%~15.61%的种子活力仍保持在60%以上;其中含水量13.58%的种子活力最高,并且玻璃化冷冻处理的种子活力(72.67%)高于快速冷冻处理的种子活力(65.81%)。据此认为,液氮超低温长期保存高良姜种子是可行的。  相似文献   

Effects of polyethylene glycol and a synthetic ice blocker during vitrification of immature porcine oocytes on survival and subsequent embryo development          下载免费PDF全文

Elisa Caroline da Silva Santos  Tamas Somfai  Ruth Appeltant  Thanh Quang Dang‐Nguyen  Junko Noguchi  Hiroyuki Kaneko  Kazuhiro Kikuchi 《Animal Science Journal》2017,88(8):1042-1048

We evaluated the effects of polyethylene glycol (PEG) and Supercool X‐1000 (SC) as supplements during the vitrification of immature cumulus‐enclosed porcine oocytes in a solution based on 17.5% ethylene glycol + 17.5% propylene glycol. After warming, the oocytes were subjected to in vitro maturation, fertilization and embryo culture. In Experiment 1, equilibration and vitrification solutions were supplemented with or without 2% (w/v) PEG (PEG+ and PEG‐, respectively). The survival rate, cleavage and blastocyst development were similar between PEG+ and PEG‐ groups; however, all values were lower than those in the non‐vitrified control. In Experiment 2, vitrification solution was supplemented with or without 1% (v/v) SC (SC+ and SC‐, respectively). The percentages of survival and blastocyst development were similar between SC+ and SC‐ groups but lower than those in the non‐vitrified control. The percentage of cleavage in the SC‐ group was significantly lower than the control and the SC+ groups, which were in turn similar to one another. In both experiments, the cell numbers in blastocysts were not significantly different among the non‐vitrified and vitrified groups. In conclusion, PEG did not improve oocyte survival and embryo development, whereas SC improved the ability of surviving oocytes to cleave but not to develop into blastocysts.  相似文献   

猪卵母细胞在GV期与MⅡ期的冷冻保存效果比较研究     

吴国权  权国波  吕春荣  邵庆勇  洪琼花 《中国畜牧兽医》2017,44(3):807-812

为比较猪卵母细胞在GV期与MⅡ期的冷冻保存效果,试验在这两个成熟阶段对其进行玻璃化冷冻,GV期卵母细胞解冻后培养至成熟,MⅡ期卵母细胞解冻后恢复2 h,然后采用免疫荧光标记、Western blotting和链霉蛋白酶溶解方法分别检测它们的皮质颗粒分布、CD9蛋白表达水平和透明带消化时间上的差异。结果表明,GV期卵母细胞在解冻后2 h的存活率显著低于MⅡ期卵母细胞（P<0.05）,但极体排出率与对照卵母细胞无明显差异（P>0.05）;在冷冻MⅡ期卵母细胞中,皮质颗粒的皮质区分布比例和CD9的蛋白表达水平显著下降（P<0.05）,但冷冻GV期卵母细胞经体外成熟后则无明显变化（P>0.05）;冷冻GV期与MⅡ期卵母细胞均不会影响透明带的消化时间（P>0.05）。由此可见,猪卵母细胞在GV期的冷冻存活率虽然较MⅡ期低,但其体外成熟后极体排出率、皮质颗粒分布和CD9蛋白表达水平均未受到冷冻的影响。  相似文献   

Cryopreservation of Queen Honeybee （Apis mellifera carnica） Born Worker Eggs by Vitrification     

LI  Zhi-yong XUE  Yun-bo WANG  Zhi LI  Xing-an 《中国农业科学(英文版)》2010,9(9):1353-1361

Many species of insect egg can be targeted individually or （and） collectively for cryopreservation by vitrification. However, there has been no report on cryopreservation of honeybee eggs by vitrification. In an attempt to define a preliminary procedure of cryopreservation of honeybee eggs by vitrification, queen honeybee born worker eggs （worker eggs） were stored through vitrification in liquid nitrogen up to 1 h, and then post-vitrification survival of the vitrified worker eggs in vitro and their hatching rates during maturation in vivo were observed using microscopic and close visual inspections. The procedure of cryopreservation by vitrification included dechorionation with sodium hypochlorite and permeabilization with isopropyl alcohol; equilibration by addition of loading solution （i.e., 25% vitrification storage solution） and dehydration by gradual replacement of loading solution with vitrification storage solution; cooling in liquid nitrogen vapor right before droplet vitrification in liquid nitrogen; and recovery in liquid nitrogen vapor right after storage in liquid nitrogen, thawing at temperature of thawing medium （5% sucrose in TC 100-insect medium） and rehydration by gradual replacement of vitrification storage solution with rehydration solution （5% fetal bovine serum in TC 100-insect medium）. It was found that among the worker eggs experiencing cyropreservation by vitrification, 1.25% of them were successfully passed through the four life stages, viz., egg, larva, pupa, and adult. In summary, it can be inferred that although a majority of worker eggs were dead after cyropreservation by vitrification, a few of them were developed into larvae, pupae, and finally emerged as adults.  相似文献   

密花石斛花粉的保存方法研究     

欧阳英  李秉玲  刘燕 《北京林业大学学报》2010,32(6):151-154

研究了密花石斛花粉不同保存方法下的花粉寿命,并对密花石斛花粉的超低温保存技术程序进行了研究。 结果表明:1)密花石斛花粉采用室温（(27±2)℃）、冷藏（4℃）、冷冻（-20℃）保存,花粉寿命分别为5、23、17 d,超低温保存(-196℃)的花粉120 d后仍可检测到花粉萌发,且萌发率与新鲜花粉相同,超低温保存方法至少可以保存密花石斛花粉4个月;2)密花石斛花粉超低温保存的程序为新鲜花粉（含水量43.26%）直接投入液氮（LN）保存,需用时采用室温、自来水或水浴化冻即可。   相似文献