Assessment of impact of varied preconditioning factors, used in isolation and combination, for enhancement of cytokine secretions in stem cell conditioned medium

ABSTRACT

Stem cell conditioned medium exhibits a huge regenerative potential but low concentration of cytokines, highdevelopment cost, and scalability challenges are major deterrents to product advances. This research studiedimpact on vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) secretions afterpreconditioning human dental pulp stem cells (HDPSCs) with diverse factors, viz., Deferoxamine (250 µM),Epidermal Growth Factor (EGF) (2 ng/ml), Angiotensin I (5mM) and Insulin Transferrin Selenium (ITS) (1%),and Niacinamide (5 mM). Additionally, advantage of using “pooled population” of HDPSCs was ascertained.HDPSCs were incubated under standard culture conditions using optimized growth media. On reaching 80%–85% confluency, media was discarded and fresh serum free media with preconditioning factors, initially oneat a time and subsequently in combination, was added. Spent media, collected after 48 hours of incubation,was used for enzyme-linked immunosorbent assay testing of selected cytokines. In comparison to control,preconditioning factors, used in isolation showed varied but significant upregulation in the secretion of VEGFbut down regulation of HGF. When used in combination, VEGF secretion increased almost 10 times of controlwith no significant change in HGF level. Results demonstrated importance of choosing right preconditioningfactors and right interplay of preconditioning factors to enhance secretions. This research paves the path todevelop an effective and commercially viable conditioned medium that can be used as “biological active” forpharmaceuticals and for developing “customized” serum free medium enriched with selected cytokines