Defined system for in vitro production of porcine embryos using a single basic medium
Defined system for in vitro production of porcine embryos using a single basic medium
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DOI：
10.1262/jrd.20001
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发表时间：
2008-06-01
影响因子：
1.8
通讯作者：
Onishi, Akira
中科院分区：
文献类型：
--
作者：
Yoshioka, Koji;Suzuki, Chie;Onishi, Akira
我们以前已经表明，猪囊胚可以通过体外受精（IVF）和培养（IVC）分别在化学成分确定的猪配子培养基（PGM）和猪受精卵培养基（PZM）-5中产生。基础培养基和大分子成分对体外成熟的影响研究了通过IVM、IVF和IVC使用单一基础培养基开发用于体外胚胎生产的确定系统。猪未成熟卵母细胞在猪卵母细胞培养基（POM）或改良的北卡罗来纳州州立大学（mNCSU）37中成熟，其补充有10%（v/v）猪卵泡液（pFF）或3 mg/ml聚乙烯醇（PVA）作为大分子组分（命名为POM+pFF、POM+PVA、mNCSU 37 +pFF和mNCSU 37 +PVA）。在mNCSU 37 +PVA组中，达到中期11期的卵母细胞比例显著低于其他处理组。在用上述培养基进行IVM后，用电刺激和放线菌酮处理卵母细胞以进行孤雌激活，并在PZM-5中培养5天。与其他处理组相比，mNCSU 37 +PVA处理组孤雌激活卵母细胞的卵裂率和囊胚形成率显著降低，而囊胚中细胞总数在处理组间差异不显著。在IVF和IVC后，当卵母细胞在mNCSU 37 +PVA中成熟时，穿透率、雄性原核形成率、卵裂率和囊胚形成率显著低于其他成熟培养基。POM+PVA的正常受精率显著高于其他处理，尽管与pFF补充相比，在POM和mNCSU 37中加入PVA会减少囊胚中的细胞总数。这些结果表明，猪囊胚可以通过使用单一基础培养基的限定系统产生。
We have previously indicated that porcine blastocysts can be produced by in vitro fertilization (IVF) and culture (IVC) in chemically defined porcine gamete medium (PGM) and porcine zygote medium (PZM)-5, respectively, In the present study, the effects of basic media and macromolecular components on in vitro maturation (IVM) were investigated to develop a defined system for in vitro embryo production using a single basic medium through IVM, IVF and IVC. Porcine immature oocytes were matured in porcine oocyte medium (POM) or modified North Carolina State University (mNCSU) 37, which were supplemented with either 10% (v/v) porcine follicular fluid (pFF) or 3 mg/ml polyvinyl alcohol (PVA) as a macromolecular component (designated POM+pFF, POM+PVA, mNCSU37+pFF and mNCSU37+PVA). In the maturation with mNCSU37+PVA, the percentages of oocytes that reached the metaphase 11 stages were significantly lower than those in the other treatments. Following IVM with the above media, oocytes were treated with an electrical stimulus and cycloheximide for parthenogenetic activation and were cultured in PZM-5 for 5 days. The rates of cleavage and blastocyst formation of parthenogenetic oocytes were significantly lowered for maturation with mNCSU37+PVA compared with the other treatments, while there were no significant differences in the total numbers of cells in blastocysts among the treatments. Following IVF and IVC, the rates of penetration, male pronucleus formation, cleavage and blastocyst formation were significantly lower when oocytes were matured in mNCSU37+PVA than in other maturation media. The normal fertilization rate was significantly higher in POM+PVA compared with the other treatments, although the total number of cells in blastocysts was reduced with the addition of PVA to both POM and mNCSU37 compared with pFF supplementation. These results demonstrate that porcine blastocysts can be produced by the defined system using a single basic medium.