Cryoprotectant formulation differentially shapes cardiac and immune transcriptomes in frozen storage

ABSTRACT: Cardiac tissue slices are used widely for biobanking and pharmacological testing, yet how freezing and cryoprotection reshape their molecular state remains poorly defined. We stored porcine cardiac tissue slices at -20 °C in a conventional DMSO-containing cryomedium or DMSO-free trehalose-based formulations, with or without supplementation, and profiled transcriptomes from day 1 to day 7 together with TUNEL apoptosis and CD68 immunophenotyping. Transcriptional injury was formulation- and time-dependent, caused by freezing rather than 4 °C handling. The DMSO-containing cryomedium drove collapse of sarcomeric, calcium-handling and mitochondrial programmes, with key transcripts falling by -3 to -4.4 log2FC at day 7. DMSO-free storage preserved these programmes substantially better. The four supplemental protectants recovered myocardial metabolic programmes and attenuated necroptotic and ferroptotic signalling. Tissue-resident immunity was the most striking finding: complement, phagocytic and NET-related transcripts were preserved at day 1 in DMSO-containing storage, fell sharply in DMSO-free storage, and partially recovered by day 7, whereas DMSO-stored tissue showed progressive immune-associated activation. Immunophenotyping revealed formulation-dependent CD68+ cell density that mirrored the transcriptomic immune signature.

ORGANISM(S): Sus scrofa

PROVIDER: GSE344759 | GEO | 2026/09/01

REPOSITORIES: GEO