Navakauskienė Rūta, Žukauskaitė Deimantė, Borutinskaitė Veronika Viktorija, Bukreieva Tetiana, Skliutė Giedrė, Valatkaitė Elvina, Zentelytė Aistė, Piešinienė Lina, Shablii Volodymyr
Department of Molecular Cell Biology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Vilnius, Lithuania.
Laboratory of Biosynthesis of Nucleic Acids, Department of Functional Genomics, Institute of Molecular Biology and Genetics, National Academy of Science, Kyiv, Ukraine.
Front Bioeng Biotechnol. 2023 Apr 13;11:1140781. doi: 10.3389/fbioe.2023.1140781. eCollection 2023.
Cryopreservation of placenta tissue for long-term storage provides the opportunity in the future to isolate mesenchymal stromal cells that could be used for cell therapy and regenerative medicine. Despite being widely used, the established cryopreservation protocols for freezing and thawing still raise concerns about their impact on molecular characteristics, such as epigenetic regulation. In our study, we compared the characteristics of human placental mesenchymal stromal cells (hPMSCs) isolated from fresh (native) and cryopreserved (cryo) placenta tissue. We assessed and compared the characteristics of native and cryo hPMSCs such as morphology, metabolic and differentiation potential, expression of cell surface markers, and transcriptome. No significant changes in immunophenotype and differentiation capacity between native and cryo cells were observed. Furthermore, we investigated the epigenetic changes and demonstrated that both native and cryo hPMSCs express only slight variations in the epigenetic profile, including miRNA levels, DNA methylation, and histone modifications. Nevertheless, transcriptome analysis defined the upregulation of early-senescence state-associated genes in hPMSCs after cryopreservation. We also evaluated the ability of hPMSCs to improve pregnancy outcomes in mouse models. Improved pregnancy outcomes in a mouse model confirmed that isolated placental cells both from native and cryo tissue have a positive effect on the restoration of the reproductive system. Still, the native hPMSCs possess better capacity (up to 66%) in comparison with cryo hPMSCs (up to 33%) to restore fertility in mice with premature ovarian failure. Our study demonstrates that placental tissue can be cryopreserved for long-term storage with the possibility to isolate mesenchymal stromal cells that retain characteristics suitable for therapeutic use.
胎盘组织的低温保存用于长期储存，为未来分离可用于细胞治疗和再生医学的间充质基质细胞提供了机会。尽管已被广泛使用，但既定的冻融低温保存方案仍引发了人们对其对分子特征（如表观遗传调控）影响的担忧。在我们的研究中，我们比较了从新鲜（天然）和低温保存（冷冻）胎盘组织中分离的人胎盘间充质基质细胞（hPMSC）的特征。我们评估并比较了天然和冷冻hPMSC的特征，如形态、代谢和分化潜能、细胞表面标志物的表达以及转录组。未观察到天然细胞和冷冻细胞之间免疫表型和分化能力的显著变化。此外，我们研究了表观遗传变化，并证明天然和冷冻hPMSC在表观遗传谱上仅表现出轻微差异，包括miRNA水平、DNA甲基化和组蛋白修饰。然而，转录组分析确定了低温保存后hPMSC中与早期衰老状态相关基因的上调。我们还评估了hPMSC改善小鼠模型妊娠结局的能力。小鼠模型中改善的妊娠结局证实，从天然和冷冻组织中分离的胎盘细胞对生殖系统的恢复具有积极作用。尽管如此，与冷冻hPMSC（高达33%）相比，天然hPMSC在恢复卵巢早衰小鼠生育能力方面具有更好的能力（高达66%）。我们的研究表明，胎盘组织可以低温保存用于长期储存，并有可能分离出保留适合治疗用途特征的间充质基质细胞。
