Araújo-Lemos Paula F B, Freitas Neto Leopoldo M, Moura Marcelo T, Melo Janaína V, Lima Paulo F, Oliveira Marcos A L
Empresa Estadual de Pesquisa Agropecuária da Paraíba (EMEPA),Rua Euripedes Tavares,210,Tambiá,CEP 58013-200,João Pessoa-PB,Brasil.
Laboratório de Biotécnicas Aplicadas a Reprodução Animal,Departamento de Medicina Veterinária,Universidade Federal Rural de Pernambuco (UFRPE),Rua Dom Manoel de Medeiros s/n,Dois Irmãos,CEP 52171-900,Recife-PE,Brasil.
Zygote. 2015 Aug;23(4):594-602. doi: 10.1017/S0967199414000215. Epub 2014 Jun 25.
The experiment aimed to compare conventional freezing and different vitrification protocols for cryopreservation of caprine embryos at morphological, ultrastructural, and functional levels. Caprine embryos produced in vivo were allocated randomly to three groups: (1) conventional freezing with ethylene glycol (EG); (2) dimethyl sulfoxide + EG (DMSO/EG) vitrification; and (3) dimethylformamide + EG (DMF/EG) vitrification. All groups were scored for cell viability (propidium iodide staining and ultrastructural levels) and re-expansion rate after thawing or warming. Embryos subjected to DMSO/EG vitrification showed higher cell viability (73.33%), compared with DMF/EG vitrification and conventional freezing group embryos (40.00 and 66.66%, respectively). The ultrastructural study revealed that vitrified embryos had greater preservation of cellular structure than embryos from conventional freezing with EG. DMSO/EG vitrification resulted in higher rates of re-expansion in vitro (47.36%) than DMF/EG vitrification (31.58%), and conventional freezing (25.00%). In conclusion, caprine embryos produced in vivo are better cryopreserved after vitrification than conventional freezing, therefore we conclude that DMSO/EG vitrification is the most effective protocol for cryopreservation.
该实验旨在从形态学、超微结构和功能水平比较常规冷冻和不同玻璃化方案对山羊胚胎进行冷冻保存的效果。将体内产生的山羊胚胎随机分为三组：（1）用乙二醇（EG）进行常规冷冻；（2）二甲基亚砜+EG（DMSO/EG）玻璃化；（3）二甲基甲酰胺+EG（DMF/EG）玻璃化。对所有组进行细胞活力评分（碘化丙啶染色和超微结构水平）以及解冻或复温后的再扩张率评估。与DMF/EG玻璃化组和常规冷冻组胚胎（分别为40.00%和66.66%）相比，接受DMSO/EG玻璃化的胚胎显示出更高的细胞活力（73.33%）。超微结构研究表明，玻璃化胚胎比用EG常规冷冻的胚胎具有更好的细胞结构保存。DMSO/EG玻璃化导致体外再扩张率（47.36%）高于DMF/EG玻璃化（31.58%）和常规冷冻（25.00%）。总之，体内产生的山羊胚胎经玻璃化冷冻保存比常规冷冻效果更好，因此我们得出结论，DMSO/EG玻璃化是最有效的冷冻保存方案。
