Birth of piglets derived from porcine zygotes cultured in a chemically defined medium
Birth of piglets derived from porcine zygotes cultured in a chemically defined medium
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DOI：
10.1095/biolreprod66.1.112
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发表时间：
2002-01-01
影响因子：
3.6
通讯作者：
Iwamura, S
中科院分区：
文献类型：
--
作者：
Yoshioka, K;Suzuki, C;Iwamura, S
我们评估了在一种新的培养基，猪受精卵培养基（PZM）中培养的猪受精卵在不同条件下的体外发育，并与体内发育进行比较。这些受精卵培养后的存活率也通过胚胎移植到受体进行了评估。在hCG注射后第2天从后备母猪中采集猪单细胞受精卵。在含3 mg/ml BSA的PZM（PZM-3）中培养的受精卵在第6天囊胚比例、第8天孵化率、第8天胚胎内细胞团（ICM）细胞数和总细胞数方面均优于北卡罗来纳州州立大学（NCSU）-23培养基。在PZM-3培养中，与空气中的5%CO2相比，在5%CO2：5%O-2：90%N-2的气氛中胚胎发育最优化。在PZM-3或其中BSA被3 mg/ml聚乙烯醇（PZM-4）替代的PZM-3中培养的第6天胚胎中的ICM和总细胞数也大于NCSU-23的ICM和总细胞数，但小于体内发育的ICM和总细胞数。然而，在PZM-3、PZM-4或体内发育的胚胎中，ICM占总细胞的比例没有差异。当将在PZM-4（99个胚胎）或体内（100个胚胎）中发育的第6天胚胎分别转移到6个受体中时，未发现产仔率（两种处理均为83.3%）和出生仔猪数量（分别为33和42头仔猪）存在差异。我们的研究结果表明，猪受精卵可以在化学成分确定的培养基中发育成囊胚，并通过培养后转移给受体而达到足月。
We evaluated the in vitro development of porcine zygotes that were cultured in a novel culture medium, porcine zygote medium (PZM), under different conditions and compared to in vivo development. The viability of these zygotes to full term after culture was also evaluated by embryo transfer to recipients. Porcine single-cell zygotes were collected from gilts on Day 2 after hCG injection. Culture of zygotes in PZM containing 3 mg/ml of BSA (PZM-3) produced better results in terms of proportion of Day 6 blastocysts, Day 8 hatching rate, and numbers of inner cell mass (ICM) cells and total cells in Day 8 embryos than that in North Carolina State University (NCSU)-23 medium. In culture with PZM-3, embryo development was optimized in an atmosphere of 5% CO2:5% O-2:90% N-2 compared to 5% CO2 in air. The ICM and total cell numbers in Day 6 embryos cultured in PZM-3 or in PZM-3 in which BSA was replaced with 3 mg/ml of polyvinyl alcohol (PZM-4) were also greater than those of NCSU-23 but less than those developed in vivo. However, no difference was found in the ratio of ICM to total cells among embryos developed in PZM-3, PZM-4, or in vivo. When the Day 6 embryos that developed in PZM-4 (99 embryos) or in vivo (100 embryos) were each transferred into six recipients, no difference was found in the farrowing rate (83.3% for both treatments) and in the number of piglets born (33 and 42 piglets, respectively). Our results indicate that porcine zygotes can develop into blastocysts in a chemically defined medium and to full term by transfer to recipients after culture.