LAQV-REQUIMTE, Campus da Caparica, Monte da Caparica, 2825-149, Caparica, Portugal.
Sci Rep. 2022 May 16;12(1):8095. doi: 10.1038/s41598-022-12365-4.
In this work we present the potential of Natural Deep Eutectic Systems (NADES) as new vitrification media for the cryopreservation of mammalian cells. Several NADES composed of natural metabolites were prepared and tested as CPAs in two cell lines, L929 and HacaT cells. After the harvesting, cells were mixed with the eutectic systems, and frozen directly into liquid nitrogen to achieve a vitreous state. Then, the cells were thawed and it was observed that NADES were able to exert a significant cryoprotective effect in L929 cells, when compared with DMSO or in the absence of a CPA. For HacaT cells, only a eutectic system showed a slightly improvement in cell survival, while DMSO caused complete cell death. Moreover, the thermal behaviour of the best systems was studied for further understanding the protective properties of NADES as CPAs, and have shown a significant difference in terms of T and T when compared with DMSO and water. Additionally, the results obtained showed that NADES can be maintained in the growth media after the thawing step, without compromising cell viability. In summary, we have shown the great potential of NADES to be used as CPAs for the cryopreservation of different cell types, using the vitrification method.
在这项工作中，我们展示了天然深共晶系统（NADES）作为哺乳动物细胞冷冻保存新的玻璃化介质的潜力。我们制备了几种由天然代谢物组成的 NADES，并将其作为两种细胞系（L929 和 HacaT 细胞）的 CPAs 进行了测试。收获细胞后，将其与共晶系统混合，并直接冷冻到液氮中以达到玻璃态。然后，将细胞解冻，结果表明与 DMSO 或没有 CPA 相比，NADES 能够显著保护 L929 细胞。对于 HacaT 细胞，只有一种共晶系统略微提高了细胞存活率，而 DMSO 则导致细胞完全死亡。此外，还研究了最佳系统的热行为，以进一步了解 NADES 作为 CPAs 的保护特性，与 DMSO 和水相比，它们在 T 和 T 方面表现出显著差异。此外，所得结果表明，NADES 可以在解冻步骤后保持在生长培养基中，而不会影响细胞活力。总之，我们已经证明 NADES 具有很大的潜力，可作为不同细胞类型的冷冻保存的 CPAs，使用玻璃化方法。
