Stem cell conditioned medium exhibits a huge regenerative potential but low concentration of cytokines, high development cost and scalability challenges are major deterrents to product advances. This research studied impact on Vascular Endothelial Growth Factor and Hepatocyte Growth Factor secretions after preconditioning human dental pulp stem cells (HDPSCs) with diverse factors viz. Deferoxamine (250 µM), EGF (2 ng/mL), Angiotensin I (5mM) and 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 one at a time and subsequently in combination, was added. Spent media, collected after 48 hours of incubation, was used for ELISA testing of selected cytokines. In comparison to control, preconditioning factors, used in isolation showed varied but significant upregulation in secretion of VEGF but down regulation of HGF. When used in combination, VEGF secretion increased almost 10 times of control with no significant change in HGF level. Results demonstrated importance of choosing right preconditioning factors and right interplay of preconditioning factors to enhance secretions. This research paves the path to develop an effective and commercially viable conditioned medium that can be used as biological active for pharmaceuticals and for developing customized serum free medium enriched with selected cytokines.
Key words: ddental pulp stem cell, conditioned medium, preconditioning factors, cytokines, VEGF, HGF
|
