Cryopreservation decreased lipopolysaccharide-induced immune response of endometrial stromal cells via inhibiting the expression of TRIF.

In the study, we investigated the effect of cryopreservation on the immune response of endometrial stromal cells (ESCs) to lipopolysaccharide (LPS). We exposed both frozen-thawed and untreated ESCs to LPS and measured cytokine levels as well as genes associated with the LPS-TLR4-cytokine signaling pathway. Results revealed that LPS induced significant upregulation of IL-1β, IL-6, IL-8, and TNF-α in vitro cultured untreated cells compared to frozen-thawed ones. Furthermore, mRNA levels related to the TRIF pathway, including TRAM, TRAF-6, and IRF-7, NF-κB, and JNK were significantly upregulated in untreated ESCs after LPS-stimulation but not in frozen-thawed cells. Moreover, transcription levels of TRIF signaling-related genes were notably lower in frozen-thawed cells. Additionally, TRIM69 overexpression rescued the cryopreservation-induced suppression of immune response. Overall, our findings suggest that cryopreservation attenuates the LPS-induced immune response in ESCs by inhibiting the TRIF pathway.