【摘要】 <正> 胚胎工程技术是动物品种、品系培育,种质资源保存及转基因动物制备、保种的重要手段。配子的冷冻保存技术目前广泛应用于胚胎工程。和胚胎冷冻相比小鼠精子冷冻技术方便、高效尤其适用于转基因及突变系小鼠的保种。成功的精子冷冻要求复苏后通过体外受精(IVF)获得胚胎,再移植入受体,生长发育并产出幼仔。胚胎体外培养获得足够的晚桑葚胚和早期囊胚是胚胎干细胞建立与制备嵌合体小鼠的成功关键,暂时不用的胚胎应仍能耐受冷冻保存,待复苏移植,建立新的繁殖群。但小鼠精子冷冻研究在我国开展的十分有限,缺乏相关资料数据。本实验对不同周龄SPF级昆明(KM)小鼠进行精子冷冻及冻融精子IVF,并将IVF获得的2-细胞胚胎分别进行胚胎移植、体外培养、胚胎冷冻及复苏后移植,应用冻融小鼠精子进行胚胎工程实践。更多还原

【Abstract】 Spermatozoa of 10, 15 and 20-wk-old KM mice were frozen and stored aat-196℃. After thawing, intact superovulation oocytes were inseminated in vitro with relative high motility frozen/thawed mouse spermatozoa. Some 2-cell embryos obtained by in vitro fertilization were transferred to pseudopregnant recipients and 47%-56% of them developed into live-born offspring. Some 2-cell embryos were incubated in vitro and 57% of them progressed to the blastocyst stage. Others were cryopreserved by simple vitrification procedure, and most of them (about 80%) were morphologically normal after warming, and 50% of the cryopreserved embryos transferred to recipients had developed into live pups. These results indicated that there was no significant difference when compared with embryos obtained from oocytes inseminated with fresh sperm.更多还原