CNRS, INSERM, Institute for Research on Cancer and Aging (IRCAN), Université Côte d'Azur, 28 avenue de Valombrose, F-06107 Nice, France.
Sorbonne Université, UFR 927, 4 Place Jussieu, F-75252 Paris, France.
Cells. 2020 Nov 26;9(12):2541. doi: 10.3390/cells9122541.
Cnidarian primary cell cultures have a strong potential to become a universal tool to assess stress-response mechanisms at the cellular level. However, primary cell cultures are time-consuming regarding their establishment and maintenance. Cryopreservation is a commonly used approach to provide stable cell stocks for experiments, but it is yet to be established for Cnidarian cell cultures. The aim of this study was therefore to design a cryopreservation protocol for primary cell cultures of the Cnidarian , using dimethyl sulfoxide (DMSO) as a cryoprotectant, enriched or not with fetal bovine serum (FBS). We determined that DMSO 5% with 25% FBS was an efficient cryosolution, resulting in 70% of post-thaw cell survival. The success of this protocol was first confirmed by a constant post-thaw survival independently of the cell culture age (up to 45 days old) and the storage period (up to 87 days). Finally, cryopreserved cells displayed a long-term recovery with a maintenance of the primary cell culture parameters and cellular functions: formation of cell aggregates, high viability and constant cell growth, and unchanged intrinsic resistance to hyperthermal stress. These results will further bring new opportunities for the scientific community interested in molecular, cellular, and biochemical aspects of cnidarian biology.
刺胞动物原代细胞培养具有成为评估细胞水平应激反应机制的通用工具的强大潜力。然而，原代细胞培养在建立和维持方面需要花费大量时间。冷冻保存是为实验提供稳定细胞库的常用方法，但尚未在刺胞动物细胞培养中建立。因此，本研究旨在设计一种使用二甲亚砜（DMSO）作为冷冻保护剂的刺胞动物原代细胞培养物的冷冻保存方案，该方案是否富含胎牛血清（FBS）。我们确定 DMSO 5%加 25% FBS 是一种有效的冷冻溶液，可使解冻后细胞存活率达到 70%。该方案的成功首先通过不受细胞培养年龄（长达 45 天）和储存期（长达 87 天）影响的恒定解冻后存活率得到证实。最后，冷冻保存的细胞具有长期恢复能力，维持原代细胞培养物的参数和细胞功能：细胞聚集的形成、高活力和持续的细胞生长，以及对过热应激的固有抗性不变。这些结果将为对刺胞动物生物学的分子、细胞和生化方面感兴趣的科学界带来新的机会。
