Sawicka Dorota, Chojnacka-Puchta Luiza, Brzezinska Jadwiga, Lakota Pawel, Bednarczyk Marek
Folia Biol (Krakow). 2015;63(2):129-34. doi: 10.3409/fb63_2.129.
Cryoconservation of blastodermal cells (BCs) can preserve genetic material for the future reconstruction of poultry breeds. The aim of our study was to compare the effects of three slow freezing programs and vitrification, different cryoprotectants (5% DMSO, 10% DMSO, or multi-component cryoprotectant (MC) and two thawing methods on the viability of chicken BCs. Significant differences in the survival of slowly frozen BCs using program 3 (2°C/min. to 0.4°C/min.) compared with programs 1 (1°C/min. to 0.3°C/min.) and 2 (4°C/min. to 0.3°C/min.) were observed. The percentage of live BCs was significantly higher after slow freezing in the presence of the MC compared with DMSO. The thawing method did not have a significant effect on the percentage of live BCs. We also observed significant differences in the survival rate of BCs after vitrification (81%) and slow freezing in the presence of 10% DMSO using program 3 (60%). The highest percentage of viable BCs was achieved by slow freezing with the MC using program 2 and thawing with method 1 (94%). The most unfavorable combination for BCs survival was slow freezing in 5% DMSO using program 3 and thawing with method 2 (58.3%). This is the first study to apply MC to the slow freezing of BCs. We also showed successful BCs vitrification.
胚盘细胞（BCs）的冷冻保存可为未来家禽品种的重建保存遗传物质。我们研究的目的是比较三种慢速冷冻程序和玻璃化、不同冷冻保护剂（5%二甲基亚砜、10%二甲基亚砜或多组分冷冻保护剂（MC））以及两种解冻方法对鸡胚盘细胞活力的影响。观察到使用程序3（2℃/分钟降至0.4℃/分钟）慢速冷冻的胚盘细胞存活率与程序1（1℃/分钟降至0.3℃/分钟）和程序2（4℃/分钟降至0.3℃/分钟）相比存在显著差异。与二甲基亚砜相比，在多组分冷冻保护剂存在下慢速冷冻后活胚盘细胞的百分比显著更高。解冻方法对活胚盘细胞的百分比没有显著影响。我们还观察到玻璃化后胚盘细胞的存活率（81%）与使用程序3在10%二甲基亚砜存在下慢速冷冻后的存活率（60%）存在显著差异。使用程序2和方法1进行多组分冷冻保护剂慢速冷冻并解冻后，获得了最高百分比的活胚盘细胞（94%）。对胚盘细胞存活最不利的组合是使用程序3在5%二甲基亚砜中慢速冷冻并使用方法2解冻（58.3%）。这是首次将多组分冷冻保护剂应用于胚盘细胞慢速冷冻的研究。我们还展示了成功的胚盘细胞玻璃化。
