Department of Obstetrics and Gynecology, Medical University of Vienna, Spitalgasse 23, 1090 Vienna, Austria.
FH Campus Wien, University of Applied Science, 1100 Vienna, Austria.
Int J Mol Sci. 2023 Jul 31;24(15):12225. doi: 10.3390/ijms241512225.
The aim of our laboratory-based study was to investigate the extent of delayed-onset cell death after cryopreservation in endothelial and epithelial cell lines of ovarian origin. We found differences in percentages of vital cells directly after warming and after cultivation for 48 to 72 h. A granulosa cell line of endothelial origin (KGN) and an epithelial cell line (OvCar-3) were used. In both DMSO-containing and DMSO-free protocols, significant differences in vitality rates between the different cell lines when using open and closed vitrification could be shown (DMSO-containing: KGN open vs. OvCar open, = 0.001; KGN closed vs. OvCar closed, = 0.001; DMSO-free: KGN open vs. OvCar open, = 0.001; KGN closed vs. OvCar closed, = 0.031). Furthermore, there was a marked difference in the percentage of vital cells immediately after warming and after cultivation for 48 to 72 h; whereas the KGN cell line showed a loss of cell viability of 41% using a DMSO-containing protocol, the OvCar-3 cell loss was only 11% after cultivation. Using a DMSO-free protocol, the percentages of late-onset cell death were 77% and 48% for KGN and OvCar-3 cells, respectively. Our data support the hypothesis that cryopreservation-induced damage is cell type and cryoprotective agent dependent.
我们的实验室基础研究旨在探究卵巢来源的内皮细胞和上皮细胞系在经过冷冻保存后迟发性细胞死亡的程度。我们发现,在解冻后直接和培养 48 至 72 小时后,活细胞的百分比存在差异。我们使用了一种内皮来源的颗粒细胞系(KGN)和一种上皮细胞系(OvCar-3)。在含有 DMSO 和不含 DMSO 的方案中,使用开放和封闭玻璃化时,不同细胞系之间的活力率存在显著差异(含有 DMSO:KGN 开放与 OvCar 开放, = 0.001;KGN 封闭与 OvCar 封闭, = 0.001;不含 DMSO:KGN 开放与 OvCar 开放, = 0.001;KGN 封闭与 OvCar 封闭, = 0.031)。此外,在解冻后立即和培养 48 至 72 小时后,活细胞的百分比有明显差异;在含有 DMSO 的方案中,KGN 细胞系的细胞活力损失为 41%,而 OvCar-3 细胞的损失仅为培养后的 11%。使用不含 DMSO 的方案,KGN 和 OvCar-3 细胞的迟发性细胞死亡百分比分别为 77%和 48%。我们的数据支持这样一种假设,即冷冻保存诱导的损伤取决于细胞类型和细胞保护剂。