Abstract

Background & Aim The regenerative and immunomodulatory properties of Mesenchymal Stem Cells (MSCs) have been demonstrated extensively in vitro, and in vivo in various types of pre-clinical models. However, in clinical settings, while the safety profile of MSCs has been well-established, the efficacy data obtained in pre-clinical studies hasn't been recapitulated in most cases where MSCs have been administered to human subjects. Many factors are thought to hinder MSC-based therapies; low potency and retention, and limited homing to the site of injury. In the current study, we sought to engineer bone-marrow-derived MSCs (BMSCs) with improved migratory properties so as to facilitate homing to the site of injury. Methods, Results & Conclusion We utilized a genetic approach in which BMSCs were transfected with CXCR4 mRNA. CXCR4, a G-protein coupled receptor highly expressed on the surface of multiple cell types is best known for its role in hematopoiesis and leukocyte chemotaxis. We showed that transfection of CXCR4 mRNA into BMSCs for eight hours resulted in over 60% of cells positive for CXCR4 protein expression, as determined by flow cytometry. Less than 1% of un-transfected BMSCs expresses CXCR4 on their surface. The expression of CXCR4 was maintained post-thaw when transfected BMSCs were recovered from LN2 Storage. Transfected cells also maintained their viability post-thaw. In terms of functionality, we demonstrated that BMSCs expressing CXCR4 were more migratory compared to their naive counterparts lacking CXCR4. This effect on migration was augmented when CXCR4-BMSCs were induced to migrate towards SDF-1α, the cognate ligand for CXCR4. In addition to the effects on directed cell migration, with the use of a membrane-based antibody array, we showed that compared to control BMSCs, CXCR4-BMSCs secrete greater quantities of MCP-1, a key chemokine that regulates macrophage migration and infiltration. In summary, we have shown that transfection of BMSCs with CXCR4 mRNA increases directed cell migration and alters cytokine secretion. This is a potential strategy for generating BMSCs with superior homing properties for use in clinical settings.

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