Effects of human placenta cryopreservation on molecular characteristics of placental mesenchymal stromal cells

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.

Automated summary

Cryopreservation of placenta tissue allows for the isolation of mesenchymal stromal cells (MSCs) for future cell therapy and regenerative medicine. However, concerns about the impact on molecular characteristics, such as epigenetic regulation, still exist. In this study, we compared the characteristics of human placental MSCs isolated from fresh and cryopreserved placenta tissue. We found that cryopreservation did not significantly affect the immunophenotype, differentiation capacity, or epigenetic profile of the MSCs. Additionally, we demonstrated that both fresh and cryopreserved MSCs from placenta tissue have a positive effect on improving pregnancy outcomes in a mouse model, with fresh MSCs showing a higher capacity to restore fertility in mice with premature ovarian failure compared to cryopreserved MSCs.