PDF(Pubmed)
Abstract：
Vitrification is a common technique for cryopreserving oocytes or embryos. However, manual vitrification is tedious and labor-intensive, and can be subject to variations caused by human factors. To address these challenges, we developed an automated vitrification-thawing system (AVTS) based on a cryo-handle. Our study firstly assessed the efficiency of cryoprotectant exchange through comparing the osmolalities of fresh and collected solutions during automated vitrification and thawing, and evaluated the cooling and warming rates of the cryo-handle. We also compared mouse oocyte survival, fertilization and embryo development after thawing and ICSI, and the development of re-frozen cleavage embryos between manual operation and automated system. The results showed that the osmolalities of collected samples were within normal range and comparable to fresh solutions. Furthermore, the automated system could obtain the reliable cooling and warming rates. Particularly, there were no significant differences in oocyte survival rates, fertilization rates, and subsequent embryo development and its quality between two procedures. Our findings suggest that AVTS has no impact on osmolalities of vitrification and thawing solutions, ensuring the proper exchange of cryoprotectants. The cryo-handle also shows the ability to achieve reliable cooling and warming rates, which benefits for the cryopreservation and thawing process. Moreover, the results from mouse oocytes and embryos indicate that automated system has effectively maintained the survival and fertilization of frozen oocytes and supported subsequent embryo development. Therefore, the automated vitrification and thawing system will inevitably represent a superior alternative to manual operation.
摘要：
玻璃化是冷冻保存卵母细胞或胚胎的常用技术。然而,手动玻璃化是繁琐和劳动密集型的，并且可能受到人为因素引起的变化。为了应对这些挑战，我们开发了一种基于冷冻手柄的自动玻璃化解冻系统（AVTS）。我们的研究首先通过比较自动玻璃化和解冻过程中新鲜和收集的溶液的渗透压来评估冷冻保护剂交换的效率，并评估了冷冻手柄的冷却和升温速率。我们还比较了小鼠卵母细胞的存活率，解冻和ICSI后的受精和胚胎发育，以及手工操作和自动化系统之间重新冷冻卵裂胚胎的发展。结果表明，收集的样品的渗透压在正常范围内，与新鲜溶液相当。此外，自动化系统可以获得可靠的冷却和升温速率。特别是,卵母细胞存活率没有显著差异，受精率,以及随后的胚胎发育及其在两个程序之间的质量。我们的研究结果表明，AVTS对玻璃化和解冻溶液的渗透压没有影响，确保冷冻保护剂的适当交换。低温处理还显示了实现可靠的冷却和升温速率的能力，这有利于冷冻保存和解冻过程。此外，小鼠卵母细胞和胚胎的结果表明，自动化系统有效地维持了冷冻卵母细胞的存活和受精，并支持了随后的胚胎发育。因此,自动玻璃化和解冻系统将不可避免地代表一个优越的替代手动操作。
