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兽医硕士香港六合资料(范文6篇)
发布时间:2019-03-20


兽医硕士香港六合资料第一篇:不同因素对小鼠冷冻精子体外受精的六合图库
 

  本篇文章目录导航:

  【题目】不同因素对小鼠冷冻精子体外受精的六合图库
  【第一章】小鼠体外受精和精子冻存研究前言
  【第二章】小鼠精子冷冻保存的材料与方法
  【第三章】体外受精和精子冻存的研究结果
  【第四章】冷冻精子不同密度对小鼠体外受精的六合图库讨论
  【第五章-参考文献】对小鼠冷冻精子和体外受精的研究结论

摘要

  
  近年来,随着小鼠品系的不断增多以及适用于不同研究需要的转基因小鼠的大量出现,使得传统的保种方式已不能满足人们的需要,精子冻存作为动物种质资源保存的一种有效手段开始被广泛应用于实验动物中,该方法具有方便快捷、成本低廉的优势而倍受研究者们的青睐,与此同时也面临着一些问题:如冷冻精子复苏后存在体外受精率不稳定、胚胎发育阻滞、产生子代的成功率低下等情况。所以本文对六合图库小鼠冷冻精子体外受精的相关因素进行研究,初步六合资料各因素对冷冻精子体外受精的六合图库机理,以期建立冻冷冻精子体外受精的最佳体系。
  
  本研究采用Nakagata精子冷冻保存方法进行小鼠精子的冷冻保存,复苏后以TYH为获能液、HTF为受精液、HTF/KSOM为早期胚胎培养液,利用控制变量的原则进行不同密度、不同获能时间、受精液中添加还原性谷胱甘肽与否、不同品系背景、不同周龄、不同基因型的小鼠冷冻精子体外受精实验,并对其早期胚胎发育结果(受精率、卵裂率、优质4-细胞胚胎率、囊胚率)进行差异显着性比较六合开奖直播,从而评价其体外受精效果。
  
  结果可见:小鼠冷冻精子密度不同,体外受精及早期胚胎发育情况差异显着,密度过低,体外受精早期胚胎发育效果极差,密度过高,受精率虽然下降幅度不大,但卵裂率较低,密度在0.8-2.0×106个/ m L范围内可以获得较好的体外受精效果;不同获能时间的冷冻精子体外受精数据显示获能时间不同,体外受精效果不同,获能时间过短(5  min)或过长(90  min)受精率、卵裂率、优质4-细胞胚胎率、囊胚率均最差,获能时间为20-40  min时,体外受精效率最高;受精液中添加还原性谷胱甘肽可显着提高冷冻精子的体外受精率及卵裂率,对优质4-细胞胚胎率及囊胚率六合图库不大;8个不同品系背景的小鼠冷冻精子体外受精早期胚胎发育情况差异显着,不同品系的小鼠精子低温生物学特性各异,杂交系和封闭群的小鼠冷冻精子体外受精效果要明显好于近交系,将其中5个品系的冷冻精子与同品系的新鲜精子进行横向比较,发现冷冻精子体外受精效率较新鲜精子均有明显下降且下降程度不成比例;不同周龄的小鼠冷冻精子体外受精结果差异亦显着,周龄过小(8周龄以下)不宜进行精子冻存及体外受精,周龄过大(30-34周)囊胚率下降幅度较大,12-30周龄小鼠冷冻精子体外受精及早期胚胎发育效果较好,其中12-24周龄最好;相同遗传背景不同基因型的小鼠冷冻精子体外受精率、卵裂率、优质4-细胞胚胎率及囊胚率也存在显着差异。
  
  综上,小鼠冷冻精子体外受精及早期胚胎发生受精子密度、获能时间、受精液中是否添加还原性谷胱甘肽、品系背景、周龄,基因型等因素的六合图库,在该冻存复苏培养体系下小鼠冷冻精子体外受精最佳密度为0.8-2.0×106个/ m L,最适获能时间为20-40 min,受精液中添加还原性谷胱甘肽可有效提高受精效率,在进行小鼠精子冷冻保种前,要注意其品系背景、周龄及基因型,以便可控的选取适宜的冷冻方法从而达到最佳复苏效果。
  
  关键字:
体外受精;小鼠;精子冻存;早期胚胎
 

  小鼠

Abstract

  
  Recently, for the reason that the strains of  mice  were increased fast, and the transgenic miceused for different research needs were appeared in mass, traditional conservation methods have beenoverwhelmed. Therefore, sperm cryopreservation has been widely used in laboratory animals whichis an effective means of animal germplasm resources conservation. The method is convenient, lowcost, which have been widely used by the researchers. However, there were still many problems inthis method, for example in the process of cryopreserved sperm in vitro fertilization after recovery,sometimes  the  fertilization  rate  is  unstable,  embryonic  development  is  blocking,  and  the  successrate of produced offspring is lower. In this paper, the factors affecting in vitro fertilization of frozensperm  in  mice  were  studied,  and  the  mechanism  of  the  factors  affecting  in  vitro  fertilization  offrozen sperm was preliminarily discussed in order to establish the best system of in vitro fertilizationof frozen sperm.
  
  In this research, Nakagata modified sperm cryopreservation methods was used to conduct theexperiments,  R18S3  was  used  as  cryopreservation  solution  for  mouse.  After  recovery,  TYH  wasused as capacitation fluid, HTF was used as fertilization medium, HTF/KSOM was used as the earlyembryo culture medium. The principles of control variables were used to conduct the fertilizationin vitro experiments of different density, different capacitation time, whether added L-GSH to thefertilization  medium  or  not,  different  strains,  different  age  and  genotypes.  The  results  of  earlyembryo development (fertilization rate, cleavage rate, high-quality 4-cell embryo rate and blastocystrate)  were  compared  and  analyzed.  The  effect  of  in  vitro  fertilization  was  evaluated,  and  themechanism of various factors was preliminarily explored in order to establish the best system of invitro fertilization of frozen-thawed sperm.
  
  Results show:
  
  The density of frozen sperm was different in mice, the development of in vitrofertilization and early embryos was significantly different. When the sperm density was too low, theeffect of in vitro fertilization was very  poor. When the sperm density is too high, the fertilizationrate decreases slightly, but the cleavage rate is low. And the density was in the range of 0.8-2.0×106个/ m L, which could obtain better in vitro fertilization results. The effect of in vitro fertilization offrozen sperm at different capacitation time is different. Fertilization rate, cleavage rate, high quality4-cell embryo rate and blastocyst rate were the worst when the capacitation time was too short (5minutes)  or  too  long  (90  minutes)。  When  the  capacitation  time  was  20-40  minutes,  in  vitrofertilization  efficiency  was  the  highest.  The  addition  of  L-GSH  in  fertilization  medium  couldsignificantly increase the fertilization rate and cleavage rate of frozen sperm in vitro, but had little effect on the rate of high-quality 4-cell embryos and blastocysts. The development of early embryosin  vitro  fertilization  of  frozen  sperm  from  8  different  strains  of  mice  was  significantly  different.
  
  Different strains of mouse sperm have different biological characteristics at low temperature.Micein hybrids and closed groups were better than inbred lines. In vitro fertilization of frozen sperm wassignificantly worse than that of fresh sperm, and the decrease was disproportionate. The results ofin  vitro  fertilization  of  frozen  sperm  in  mice  of  different  age  were  also  different.  Spermcryopreservation  and  in  vitro  fertilization  are  not  suitable  for  mice  under  8  weeks  of  age.  Theblastocyst  rate  of  mice  over  30-34  weeks  of  age  decreases  greatly.  The  effect  of  frozen  sperm  invitro fertilization and early embryo development is better in mice aged 12-30 weeks. Among them12-24  weeks  is  best.  The  in  vitro  fertilization  rate,  cleavage  rate,  high  quality  4-cell  embryo  rateand  blastocyst  rate  of  frozen-thawed  sperm  of  different  genotypes  of  mice  were  significantlydifferent.
  
  In summary, the in vitro fertilization of frozen sperm and early embryo development of mouseaffected  by  sperm  density,  capacitation  time,  whether  L-GSH  was  added  to  fertilization  medium,strain background, age, genotype and other factors, The optimum density of in vitro fertilization offrozen  sperm  was  0.8-2.0×106个/m L, and the optimum capacitation time is 20 min-40 min. Theaddition of L-GSH in fertilization medium can effectively improve fertilization efficiency. In orderto  select  the  appropriate  freezing  method  to  achieve  the  best  recovery  effect,  attention  should  bepaid to its strain background, age and genotype before cryopreservation of mouse sperm.
  
  Key words:In vitro fertilization; Mouse; Cryopreservation of spermatozoa; Early embryo
 

目录

  摘要
  英文摘要

  1前言
  1.1体外受精
  1.1.1体外受精概述
  1.1.2体外受精的应用
  1.1.3体外受精的六合图库因素
  1.1.4体外受精的研究进展

  1.2精子冷冻保存
  1.2.1精子冻存概述
  1.2.2冷冻原理和冷冻对精子功能的六合图库
  1.2.3精子冻存方法
  1.2.4精子冻存的应用
  1.2.5精子冻存的研究进展
  1.3研究目的与意义

  2材料与方法
  2.1材料
  2.1.1实验动物与样本
  2.1.2实验试剂
  2.1.3主要仪器与设备
  2.1.4主要培养液的配制

  2.2试验方法
  2.2.1小鼠精子冷冻保存
  2.2.2冻存精子复苏
  2.2.3体外受精及早期胚胎体外培养
  2.2.4受精卵及早期胚胎质量评估
  2.2.5实验设计
  2.2.6实验数据统计六合开奖直播

  3结果
  3.1体外受精及早期胚胎发育的形态学观察
  3.1.1冷冻精子、体外受精及受精卵的观察
  3.1.2早期胚胎发育的观察
  3.2不同密度的小鼠冷冻精子体外受精

  3.3不同获能时间的小鼠冷冻精子体外受精
  3.4受精液中添加还原性谷胱甘肽的小鼠冷冻精子体外受精
  3.5不同品系背景的小鼠冷冻精子体外受精
  3.5.1不同品系背景小鼠冷冻精子之间的比较
  3.5.2  不同品系背景小鼠冷冻精子与同品系新鲜精子之间的比较

  3.6  不同周龄的小鼠冷冻精子体外受精
  3.7  不同基因型的小鼠冷冻精子体外受精

  4  讨论
  4.1  冷冻精子不同密度对小鼠体外受精的六合图库
  4.2  不同获能时间对小鼠冷冻精子体外受精的六合图库
  4.3  受精液中添加还原性谷胱甘肽对小鼠冷冻精子体外受精的六合图库

  4.4  不同品系背景对小鼠冷冻精子体外受精的六合图库
  4.5  不同周龄对小鼠冷冻精子体外受精的六合图库
  4.6  不同基因型对小鼠冷冻精子体外受精的六合图库
  5  结论
  致谢

 
兽医硕士香港六合资料(优秀范文6篇)
第一篇:不同因素对小鼠冷冻精子体外受精的六合图库 第二篇:灭活益生菌静脉注射对小鼠免疫功能六合图库研究
第三篇:胚胎源外泌体对牛克隆胚胎体外发育六合图库 第四篇:山银花黄芩提取物对肉鸡生长的六合图库
第五篇:鸭细小病毒病灭活疫苗的研究 第六篇:银黄二陈合剂制备工艺和质量标准的研究

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