(150w) Unraveling the Enhancing Effect in Proliferation of Human Mesenchymal Stem Cells Cultured on Biopolymeric Coatings

Stem cell therapy is a type of tissue regeneration that attempts to repair damaged body cells by reducing inflammation and adjusting the immune system. To improve human stem cell capacity to modulate the immune system, a bioactive surface made of heterologous heparin and collagen was used. In this study, we examined viability and proliferation of hMSCs on different types of coatings with and without the FGF-2 inhibitor drug AZD4547 at a concentration of 1 µM in the culture medium. Six different types of surfaces were analyzed, including a control surface (TCPS), a surface of 6 layers of HEP/COL, and 6.5 layers of HEP/COL with and without the FGF-2 inhibitor drug AZD 4547. hMSCs were seeded on each surface of a 96-well plate, and after 0, 1, 2, and 3 days of culture, For the viability of hMSCs, PrestoBlue's method was used. The cell culture medium was removed and replaced with 100 µL of PrestoBlue reagent per well and an incubation period of 3 hours to detect fluorescence. For the proliferation of hMSCs on COL/HEP coatings, detecting cells with EdU, 18 hours is the period of EdU exposure to hMSCs. Then, cells were fixed and permeabilized, EdU was detected, DNA was stained, and finally, images and data were analyzed. HEP/COL multilayers can increase hMSCs viability compared to the control surface. After 3 days, the results showed that the viability of hMSC was higher than that of TCPS, especially on surfaces with 6.5 heparin and collagen. Furthermore, when hMSCs are grown in COL/HEP coatings, their proliferation increases. Inhibition studies suggest that HEP/COL coatings may increase hMSC proliferation through activation of the FGF-2 pathway.