Production of piglets derived from in vitro-produced Blastocysts fertilized and cultured in chemically defined media: Effects of theophylline, adenosine, and cysteine during in vitro fertilization
Production of piglets derived from in vitro-produced Blastocysts fertilized and cultured in chemically defined media: Effects of theophylline, adenosine, and cysteine during in vitro fertilization
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DOI：
10.1095/biolreprod.103.020081
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发表时间：
2003-12-01
影响因子：
3.6
通讯作者：
Rodriguez-Martinez, H
中科院分区：
文献类型：
--
作者：
Yoshioka, K;Suzuki, C;Rodriguez-Martinez, H
为了进一步开发体外成熟猪卵母细胞的体外受精（IVF）和体外培养（IVC）的条件，我们评估了茶碱，腺苷和半胱氨酸在IVF过程中的化学成分确定的培养基中的作用。还通过胚胎移植到受体来研究在化学成分确定的培养基中的IVF和IVC后体外产生的囊胚的足月存活率。猪卵母细胞培养基（PGM）是由猪受精卵培养基（PZM-4）改良而成的一种化学成分确定的培养基。PGM用作IVF的基础培养基，PZM-4用于推定合子的培养。添加2.5 mM茶碱的PGM显着增加了男性原核形成的百分比与对照组相比（无添加）。添加1 μ M腺苷的PGM补充或不补充2.5 mM茶碱显着减少穿透精子的数量与对照组相比（不添加腺苷）。与不补充半胱氨酸相比，在含有2.5 mM茶碱和1 μ M腺苷的PGM中补充0.2 μ M半胱氨酸进一步增加了发育到胚泡阶段的百分比，但无论茶碱和腺苷的存在与否，受精参数（如单精子和原核形成）均无差异。当第5天囊胚转移到4个受体（每个受体20-25个囊胚）时，所有受体都怀孕并分娩了总共21头活仔猪。目前的结果清楚地表明，猪囊胚可以产生的IVF和IVC在化学成分确定的培养基，它们可以发育到足月胚胎移植后。
To further develop defined conditions for in vitro fertilization (IVF) and in vitro culture (IVC) of in vitro-matured porcine oocytes, we evaluated the effects of theophylline, adenosine, and cysteine in a chemically defined medium during IVF. Viability to full term of in vitro-produced blastocysts after lVF and IVC in chemically defined medium was also investigated by embryo transfer to recipients. A chemically defined medium, porcine gamate medium (PGM), was modified from porcine zygote medium (PZM-4), which was previously established. PGM was used as a basal medium for lVF and PZM-4 was for the culture of presumptive zygotes. Addition of 2.5 mM theophylline to PGM significantly increased the percentage of male pronuclear formation compared with controls (no addition). Addition of 1 muM adenosine to PGM supplemented either with or without 2.5 mM theophylline significantly reduced the number of penetrated spermatozoa compared with controls (no addition of adenosine). Supplementation with 0.2 muM cysteine in PGM containing both 2.5 mM theophylline and 1 muM adenosine further increased the percentage of development to the blastocyst stage, compared with no supplementation of cysteine, but there was no difference in fertilization parameters, such as monospermy and pronuclear formation, regardless of presence or absence of theophylline and adenosine. When Day 5 blastocysts were transferred into four recipients (20-25 blastocysts per recipient), all recipients became pregnant and farrowed a total of 21 live piglets. The present results clearly demonstrate that porcine blastocysts can be produced by lVF and IVC in chemically defined media and that they can develop to full term after embryo transfer.