Piglets born after vitrification of embryos using the open pulled straw method
Piglets born after vitrification of embryos using the open pulled straw method
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DOI：
10.1006/cryo.2000.2273
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发表时间：
2000-09-01
期刊：
影响因子：
2.7
通讯作者：
Terqui, M
中科院分区：
文献类型：
--
作者：
Berthelot, F;Martinat-Botté, F;Terqui, M
以大白色多产（LWh）和梅山（MS）的桑椹胚和未孵化囊胚为材料，采用两种玻璃化冷冻液对玻璃化冷冻胚胎进行了研究。在实验1中，比较了两种冷冻保护剂稀释培养基用于冷冻保存MS和LWh囊胚：TCM是标准Hepes缓冲的TCM 199 + 20%NBCS培养基，PBS是PBS +20%NBCS培养基。在乙二醇、二甲基亚砜和蔗糖中进行两步平衡后，将2-5个囊胚装入OPS中并投入液氮中。使胚胎升温：应用具有降低浓度的蔗糖的四步稀释。在PBS中，LWh囊胚（27%）的体外活力低于MS囊胚（67%; P = 0.001）。TCM中，基因型之间没有观察到显著差异（LWh为41%，MS为43%），两种囊胚的存活率均低于对照组。在实验2中，桑椹胚阶段LWh和MS胚胎玻璃化并使用PBS加温。存活率较低，LWh（11%）和MS（14%）之间没有差异。在实验3中，将200个MS和200个LWh囊胚如实验1（PBS）中所述玻璃化/加温。在20个MS受体中，每个受体移植20个胚胎。产仔率为55%，受体分别为MS和LWh囊胚产仔4头和5头仔猪（中位数）。因此，OPS方法适用于未孵化猪囊胚的冷冻保存。(C)北京大学出版社.
Morulae and unhatched blastocysts from Large White hyperprolific (LWh) and Meishan (MS) gifts were selected to test an ultrarapid open pulled straw (OPS) vitrification method with two media, The viability of vitrified/warmed embryos was estimated by the percentage of embryos that developed to the hatched blastocyst stage in vitro or by birth after transfer. In Experiment 1, two cryoprotectant dilution media were compared for cryopreservation of MS and LWh blastocysts: TCM was a standard Hepes-buffered TCM199 + 20% NBCS medium and PBS was a PBS + 20% NBCS medium. After a two-step equilibration in ethylene glycol, dimethyl sulfoxide, and sucrose, 2-5 blastocysts were loaded into OPS and plunged into liquid nitrogen. Embryos were warmed: a four-step dilution with decreasing concentrations of sucrose was applied. In PBS, LWh blastocysts (27%) had a lower viability in vitro than MS blastocysts (67%; P = 0.001). In TCM, no significant difference was observed between genotypes (41% for LWh and 43% for MS blastocysts) and both viability rates were lower than that of the control groups. In Experiment 2, morula-stage LWh and MS embryos were vitrified and warmed using PBS. The viability rate was low and did not differ between LWh (11%) and MS (14%). In Experiment 3, 200 MS and 200 LWh blastocysts were vitrified/warmed as described in Experiment 1 (PBS). In each of 20 MS recipients, 20 embryos were transferred. The farrowing rate was 55% and recipients farrowed four and five piglets (median) for MS and LWh blastocysts, respectively. The OPS method is therefore appropriate for cryopreservation of unhatched porcine blastocysts. (C) 2000 Academic Press.