Whole transcriptome RNA sequencing analysis on mesenchymal stem cells preconditioned by inflammatory stimuli

Background & Aim Mesenchymal stem cells are a promising cell type in the field of cell therapy, and the number of clinical trials utilizing different tissue sources derived MSCs continues to grow. To date, various cell preconditioning strategies have been used to boost MSC's existing immunomodulatory functions and include exposure to environmental stimuli or small molecules. However, the exact molecular mechanism by which preconditioning enhances MSCs potency, particularly with inflammatory stimuli, is still not well understood. Herein we performed whole-transcriptome RNA sequencing (RNAseq) on cytomix preconditioned MSCs (to mimic the acute inflammatory microenvironment) to specifically examine MSC molecular pathways involved by this preconditioning strategy. Methods, Results & Conclusion Human bone marrow MSCs were used between passages 3-6, and MSCs with or without cytomix preconditioning were collected for whole transcriptome RNAseq (Qiagen). Analysis of the transcriptome profile showed a robust separation of gene expression profiles for cytomix-preconditioned versus non-preconditioned MSCs. Differential expression analysis (DESEq2) revealed 1,020 upregulated genes (FDR 1) and 413 downregulated genes (FDR In summary, our preliminary analysis on the whole transcriptome of inflammatory stimuli-preconditioned MSCs showed upregulation of genes coding for secreted proteins associated with functions in immune responses. Further experiments will be performed to confirm the relationships between genes coding for secreted proteins identified in this study with MSC's immunomodulatory functions.