Abstract: Objective To investigate the dynamic transcriptomic changes of Chinese hamster ovary (CHO) cells during reduced-temperature culture and to characterize the temporal patterns and potential regulatory features of gene expression under low-temperature conditions. Methods CHO cells were cultured under standard conditions (37 ℃) and reduced temperature (30 ℃) separately. Cells were harvested at 0 h, 24 h, and 48 h for RNA sequencing (RNA-seq). Both preprocessing and downstream analyses of the sequencing data were performed, including differential gene expression analysis, temporal expression clustering, functional enrichment. Protein-protein interaction (PPI) network analysis was performed to evaluate alterations in key signaling pathways during temperature reduction. Results Temperature reduction markedly altered the transcriptomic profile of CHO cells. Temporal clustering identified six distinct gene expression patterns, mainly associated with protein translation and ribosome biogenesis, cell cycle regulation, and vesicle transport. Biomolecular interaction network analysis revealed that translation-related molecules and membrane structure/signaling-related nodes formed relatively concentrated interaction modules, exhibiting differential temporal dynamics. Conclusions CHO cells undergo pronounced time-dependent transcriptomic remodeling during reduced-temperature culture characterized by stage-specific adjustments in translation-related and membrane structure-related processes. These findings provide transcriptome-level evidence for cellular state transitions and production-related regulation in low-temperature.

Key words: Chinese hamster ovary(CHO) cell, temperature downshift, transcriptomics, time-series analysis, biomolecular interaction network analysis