High-throughput cryopreservation of in vivo-derived swine embryos.
High-throughput cryopreservation of in vivo-derived swine embryos.
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DOI：
10.1371/journal.pone.0065545
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Prather RS
中科院分区：
文献类型：
--
作者：
Spate LD;Murphy CN;Prather RS
猪胚胎的冷冻保存效率低下。我们的目标是开发一种非侵入性方法，对体内产生的猪胚胎进行“相对”高通量的冷冻保存。由于去除早期猪胚胎中的脂滴可以改善冷冻存活率，因此我们希望将首先为体外产生的猪胚胎开发的高渗透压处理和离心技术应用到体内产生的猪胚胎。第一个目的是确定体内产生的受精卵和 2 细胞阶段胚胎在短时间内对各种高渗透压条件的敏感性。在 300、400 或 500 毫渗摩尔 (mOsm) 下培养 6、12 或 18 分钟对所得囊胚阶段胚胎没有可检测到的影响（内细胞团核数、滋养外胚层核、细胞核总数、滋养外胚层与内细胞团核的比率或囊胚百分比）。然而，后备母猪对这些参数中的每一个都有影响。对于第二个目标，我们专注于 300 mOsm 6 分钟、400 mOsm 12 分钟、500 mOsm 12 分钟和 500 mOsm 18 分钟。在高渗透压处理期间对胚胎进行离心，然后培养至囊胚阶段并玻璃化。玻璃化和解冻后，500 mOsm 18 分钟的再膨胀百分比最高，细胞核总数没有差异。虽然需要与体外产生的胚胎不同的基础培养基，但体内衍生的胚胎也能在冷冻保存中存活，而不会损害其透明带。
Cryopreservation of swine embryos is inefficient. Our goal was to develop a non-invasive method for “relatively” high-throughput cryopreservation of in vivo-produced swine embryos. Since removal of the lipid droplets within early swine embryos improves cryosurvival we wanted to apply a technique of high osmolality treatment followed by centrifugation that was first developed for in vitro-produced swine embryos to in vivo-produced swine embryos. The first aim was to determine how sensitive the in vivo-produced zygote and 2-cell stage embryo was to various high osmolality conditions for a short duration. Culture for 6, 12 or 18 min at 300, 400 or 500 milliosmoles (mOsm) had no detectable affect on the resulting blastocyst stage embryos (number of inner cell mass nuclei, trophectoderm nuclei, total number of nuclei, ratio of the trophectoderm to inner cell mass nuclei or percent blastocyst). However there was an effect of gilt on each of these parameters. For the second aim we focused on 300 mOsm for 6 min, 400 mOsm for 12 min, 500 mOsm for 12 min, and 500 mOsm for 18 min. The embryos were centrifuged for the duration of high osmolality treatment, then cultured to the blastocyst stage and vitrified. After vitrification and thawing the 500 mOsm for 18 min had the highest percent re-expansion with no difference in the total number of nuclei. While requiring a different base culture medium than in vitro-produced embryos, in vivo-derived embryos also survive cryopreservation without damage to their zona pellucida.
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影响因子：
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通讯作者：
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