CINPIC, Fish Culture Research Institute, School of Veterinary Medicine and Zootechnics, Department of Aquaculture Sciences, University of Córdoba, Cra 6, 77-305, Montería 230002, Colombia.
School of Basic Sciences, Department of Biology, University of Córdoba, Cra 6, Montería 77-305, Colombia.
Cells. 2023 Feb 9;12(4):562. doi: 10.3390/cells12040562.
Each phytoplankton species presents a different behavior and tolerance to the cryopreservation process. Therefore, in a species-specific protocol, it is essential to ensure both growth and post-thawing cell viability. In this study, we explored the effect of cryopreservation of sp. with two cryoprotectants, dimethyl sulfoxide (DMSO) and methanol (MET), at 5% and 10% inclusion for each. In the control treatment, the microalgae were not exposed to cryoprotective agents (Control). Three post-thawing cell viability criteria were used: no cell damage (NCD), cell damage (CD), and marked lesions (LM), and mitochondrial and cell membrane damage was evaluated by flow cytometry. The study was a 2 × 2 factorial design, with five replications by treatments, population growth, and cell damage evaluated from the fifth day after thawing. On the fifth day, the highest percentage of NCD was observed when the microalgae were cryopreserved with DMSO 5% (50%); Regarding the control group, it showed 0% NCD. Flow cytometry analysis reveals minor damage at the membrane and mitochondria (9-10.7%) when DMSO is used at both inclusion percentages (5-10%) after thawing. In the exponential phase, the highest growth rates, doubling time, and yield was observed in cryopreserved cells with MET 5%. The results suggest that DMSO 5% is an ideal treatment for cryopreserving microalgae sp.
每种浮游植物在冷冻保存过程中表现出不同的行为和耐受性。因此，在特定于物种的方案中，确保生长和冻后细胞活力是至关重要的。在这项研究中，我们探讨了用两种冷冻保护剂二甲亚砜（DMSO）和甲醇（MET）在 5%和 10%的包含物水平下冷冻保存 sp.的效果。在对照处理中，微藻未暴露于冷冻保护剂（对照）。使用三种冻后细胞活力标准：无细胞损伤（NCD）、细胞损伤（CD）和明显损伤（LM），并通过流式细胞术评估线粒体和细胞膜损伤。该研究采用 2×2 析因设计，每个处理有五个重复，从解冻后第五天开始评估种群生长和细胞损伤。在第五天，当用 5% DMSO 冷冻保存微藻时，观察到最高的 NCD 百分比（50%）；关于对照组，它显示出 0%的 NCD。在解冻后使用 DMSO 的两种包含物百分比（5-10%）时，流式细胞术分析显示细胞膜和线粒体有轻微损伤（9-10.7%）。在指数期，用 MET 5%冷冻保存的细胞观察到最高的生长率、倍增时间和产率。结果表明，5% DMSO 是冷冻保存 sp.的理想处理方法。
