The Effect of Lipopolysaccharide on TSG6 Expression in Mesenchymal Stromal Cells

Abstract

Purpose : Mesenchymal stem cells (MSCs) play an important role during inflammation and its resolution. We evaluated the effect of lipopolysaccharides (LPS) found in gram-negative bacteria on TSG6 expression (a well-known anti-inflammatory agent) in the corneal-limbal and bone-marrow derived MSCs (CL-MSC and BM-MSC).

Methods : BM-MSC were isolated from human bone marrow donors in a GMP facility. CL-MSCs were extracted from healthy cadaver corneas. MSCs were grown to confluency using MEM-alpha + 10% fetal bovine serum. Telomerase-immortalized human corneal epithelial cells (HCEC) were used as an external control to MSCs. They were stimulated with 100 ng/ml LPS for 24 hours, followed by mRNA extraction. After reverse transcription, the mRNA expression of TSG6 was analyzed with ^delta-deltaCt method by real-time qPCR using GAPDH as control.

Results : LPS treatment resulted in a 4.7 and 5.78 folds increase in TSG6 mRNA expression in CL-MSCs and BM-MSCs compared to the unstimulated cells, respectively (P < 0.01). There were no significant changes in TSG6 mRNA expression in HCECs after incubation with LPS.

Conclusions : LPS exposure leads to upregulation of TSG6 in MSCs. This response is observed in MSCs and not HCECs. Protein expression is needed to confirm these results.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.