Enhancing cryopreservation of human iPSCs: Bottom-up vs Conventional freezing geometry

Teodoro Duarte Garcia Morais, Fernando Jorge; El-Guendouz, Soukaina; Neves, Rafaela; Duarte, Andreia; Rodrigues, Miguel A.; Pinho Melo, Eduardo

http://hdl.handle.net/10400.1/26786

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Authors

Teodoro Duarte Garcia Morais, Fernando Jorge

El-Guendouz, Soukaina

Abstract(s)

Induced pluripotent stem cells (iPSCs) hold large potential on regenerative medicine due to their pluripotency and unlimited self-renewal capacity without the ethical issues of embryonic stem cells. To provide quality-controlled iPSCs for clinical therapies, it is essential to develop safe cryopreservation protocols for long-term storage, preferable amenable for scale-up and automation. We have compared the impact of two different freezing geometries (bottom-up and conventional radial freezing) on the viability and differentiation capability of human iPSCs. Our results demonstrate that the bottom-up freezing under optimized conditions significantly increases iPSCs viability, up to 9% for the cell membrane integrity and up to 21% for the cell metabolic state, compared to conventional freezing. The improvement achieved for bottom-up versus conventional freezing was maintained after scale-up from cryogenic vials to 30 mL bags, highlighting the method’s potential for clinical applications. These findings show that bottom-up freezing can offer a more controlled and scalable cryopreservation strategy for iPSCs, promoting their future use in regenerative medicine.