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Percoll法处理牛精子对体外受精胚胎发育的影响 被引量:13

Effect of Bovine Sperm Separation by Percoll Method on Development of Embryos Fertilized in Vitro
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摘要 牛冷冻精液解冻后,在Percol梯度液中400g离心24min,有效地获得了57.87%±8.0%的精子回收率;对照组上游法精子回收率仅为15.87%±1.9%(P<0.05)。Percol和上游2种方法分离的精子在受精液中8、18h后的活率,分别为60%、10%和70%、20%。2种浓度Percol法分离的精子对不同成熟处理的卵母细胞体外受精(IVF)卵裂率的影响:精子最终浓度为2×106/mL时,成熟组Ⅰ(LH+FSH)和成熟组Ⅱ(PMSG+hCG)的卵裂率分别为56.19%和58.16%;精子最终浓度为5×106/mL时,2组的卵裂率分别为59.30%和62.77%,卵裂率在精子浓度不同组间及添加不同激素组间的差异不显著。本试验还研究了2种方法处理的精子对不同成熟处理组的卵裂率和囊胚率,用Percol法处理的精子(2×106/mL)在成熟组Ⅰ获得了57.16%的卵裂率和25.58%的囊胚率,而上游法分别为61.95%和19.02%,2种方法相差不显著(P>0.05);在成熟组Ⅱ获得了62.18%的卵裂率和24.77%的囊胚率,而上游法分别为71.18%和20.96%,2种方法的卵裂率相差显著(P<0.? The study was designed for 4 experiments:(1) The centrifugation of the semen on the Percoll gradient treatment at 400 g for 24 min was sufficient to obtain optimal recovery of the motile spermatozoa with the recovery rate of Percoll and swim up treatment being 57.87%±8% and 15.87%±1.9%(P<0.05) respectively. (2) The sperm motility that separated by Percoll and swim up methods at 8 and 18 h in Fert TALP and 2×10 6/mL concentration were 60%, 10% and 70%, 20% respectively. (3) Two concentration of spermatozoa separated by Percoll were added to two maturation groups of oocyte, groupⅠ was treated by LH+FSH and groupⅡ by PMSG+hCG with the cleavage rates at 2×10 6/mL and 5×10 6/mL being 56.19%, 58.16%(P>0.05) and 59.3%, 62.77%(P> 0.05 ) respectively. (4) At 2×10 6/mL spermatozoa concentration, by Percoll and swim up methods, the cleavage rate and blastocyst rate in group Ⅰ were 57.16%, 61.95%(P>0.05) and 25.58 %, 19.02% (P>0.05), in groupⅡ were 62.18%, 71.18% and 24.77%, 20.96% respectively. (5) The cell number of blastocyst, by Percoll and swim up methods were 63.79%±11.34% and 67.17 %±14.81%(P>0.05). These data indicated that there were same development ability in the bovine embryos treated by Percoll and swim up separation procedure and that the Percoll method was simpler, faster, and recovered more motile spermatozoa than that of swim up. It is possible to obtain clean semen prepared for IVF using Percoll method.
出处 《中国兽医学报》 CAS CSCD 北大核心 1998年第4期394-397,共4页 Chinese Journal of Veterinary Science
关键词 Percoll法 冷冻精子 体外受精 胚胎 bovine Percoll method frozen semen in vitro fertilization embryo
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