Department of Intracellular Signaling and Transport, Institute of Cytology, Russian Academy of Sciences, Tikhoretskii pr. 4, St. Petersburg 194064, Russia.
Int J Mol Sci. 2024 Jan 25;25(3):1467. doi: 10.3390/ijms25031467.
Cryopreservation is an essential step for utilizing various cell types for biological research and medical purposes. At the same time, there is a lack of data on the effect of cryopreservation, especially when prolonged, on the karyotype of cells. In the present work, we analyzed the genetic stability of cells subjected to a cryopreservation procedure. The objects were immortalized Chinese hamster lung fibroblasts (CHL V-79 RJK line) and human endometrial mesenchymal stem/stromal cells (eMSCs). We showed that short-term cryopreservation in liquid nitrogen for up to 6 months did not affect the karyotype stability of CHL V-79 RJK and eMSCs. On the contrary, karyotyping of G-banded metaphase chromosomes in cells underwent 10-year cryopreservation, which revealed genomic instability in both cell lines associated with the variability of chromosome number in cells, random chromosomal rearrangements, and condensation disorder in homologs. In addition, we found out that long-term cryopreservation of eMSCs does not affect the expression of their typical surface markers and morphology, but results in a significant reduction in proliferative potential and early manifestation of cellular senescence features upon eMSCs culturing. Thus, we concluded that the long-term cryopreservation of cells of different types and biological origin can lead to irreversible changes of their karyotype and acceleration of cellular senescence.
冷冻保存对于利用各种细胞类型进行生物研究和医学用途至关重要。同时，关于冷冻保存对细胞染色体组型的影响，特别是长期冷冻保存的影响，数据仍然缺乏。在本工作中，我们分析了经过冷冻保存程序的细胞的遗传稳定性。研究对象是永生的中国仓鼠肺成纤维细胞（CHL V-79 RJK 系）和人子宫内膜间充质干细胞（eMSCs）。结果表明，在液氮中短期冷冻保存长达 6 个月不会影响 CHL V-79 RJK 和 eMSCs 的染色体组型稳定性。相反，对经过 10 年冷冻保存的细胞进行 G 带中期染色体核型分析，揭示了两条细胞系的基因组不稳定性，与细胞数量的变异性、随机染色体重排以及同源染色体的凝聚紊乱有关。此外，我们发现 eMSCs 的长期冷冻保存不会影响其典型表面标志物和形态的表达，但会导致其增殖潜力显著降低，并在培养 eMSCs 时早期表现出细胞衰老特征。因此，我们得出结论，不同类型和来源的细胞的长期冷冻保存可能导致其染色体组型发生不可逆转的变化，并加速细胞衰老。
