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高渗介质处理在胚胎显微操作中的潜力:I. 蔗糖处理后兔胚胎在体外和体内的存活情况

Potential of hypertonic medium treatment for embryo micromanipulation: I. Survival of rabbit embryos in vitro and in vivo following sucrose treatment.

作者信息

Yang X, Chen Y, Chen J, Foote R H

机构信息

Department of Animal Science, Cornell University, Ithaca, New York 14853-4801.

出版信息

Mol Reprod Dev. 1990 Oct;27(2):110-7. doi: 10.1002/mrd.1080270205.

Abstract

Rabbit zygotes and embryos were exposed to hypertonic sucrose in phosphate-buffered saline (SPBS). In experiment one, 144 zygotes shrank to 32-36% of their initial volume in 1.0 M SPBS within 30 min. Neither hypertonic treatment with 0.5 M or 1.0 M SPBS nor micropuncture of the zona pellucida after shrinkage affected embryo development into blastocysts in vitro (88%, 83%, and 82%, respectively), compared to that of the controls (93%, P greater than .05). In experiment two, 252 two- to four-cell- and 177 morula-stage embryos were exposed to isotonic PBS control or 0.5 M, 1.0 M, or 1.5 M SPBS for 30, 60, 90, 120, and 150 min before transfer to PBS (290 mOsm). Embryo development was significantly reduced (P less than .05) when embryos were exposed in 0.5 M and 1.0 M SPBS for more than 60 min or in 1.5 M SPBS for more than 30 min. In experiment 3, morulae exposed for 60 min to 0.5 M or 1.0 M SPBS shrank to 37-39% or 32-35% of their initial volume and then expanded to 87-94% or 81-90% of their initial volume, respectively, after being returned to isotonic PBS for 60 min, but embryos in 1.5 M SPBS had erratic osmotic behavior. In experiment four, 192 two- to four-cell embryos exposed to 0.5 M SPBS for 0, 30, and 60 min before transfer to oviducts of recipients resulted in the production of 39%, 42% and 31% young, respectively (P greater than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

将兔受精卵和胚胎置于磷酸盐缓冲盐溶液(SPBS)中的高渗蔗糖溶液中。在实验一中,144个受精卵在1.0 M SPBS中30分钟内体积缩小至初始体积的32 - 36%。与对照组(93%)相比,用0.5 M或1.0 M SPBS进行高渗处理以及体积缩小后对透明带进行微穿刺均不影响胚胎在体外发育成囊胚(分别为88%、83%和82%,P大于0.05)。在实验二中,252个二至四细胞期胚胎和177个桑椹胚期胚胎在转移至PBS(290 mOsm)前,分别在等渗PBS对照液或0.5 M、1.0 M或1.5 M SPBS中暴露30、60、90、120和150分钟。当胚胎在0.5 M和1.0 M SPBS中暴露超过60分钟或在1.5 M SPBS中暴露超过30分钟时,胚胎发育显著降低(P小于0.05)。在实验三中,暴露于0.5 M或1.0 M SPBS 60分钟的桑椹胚体积分别缩小至初始体积的37 - 39%或32 - 35%,然后在回到等渗PBS 60分钟后分别扩展至初始体积的87 - 94%或81 - 90%,但在1.5 M SPBS中的胚胎渗透行为不稳定。在实验四中,192个二至四细胞期胚胎在转移至受体输卵管前分别在0.5 M SPBS中暴露0、30和60分钟,幼仔产出率分别为39%、42%和31%(P大于0.05)。(摘要截短于250字)

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