Haseli, MahsaCastilla-Casadiego, David A.Pinzon-Herrera, LuisHillsley, AlexanderMiranda-Munoz, Katherine A.Sivaraman, SrikanthRosales, Adrianne M.Rao, Raj R.Almodovar, Jorge2024-03-122024-03-122021-12-23Haseli, Mahsa, David A. Castilla-Casadiego, Luis Pinzon-Herrera, Alexander Hillsley, Katherine A. Miranda-Munoz, Srikanth Sivaraman, Adrianne M. Rosales, Raj R. Rao, and Jorge Almodovar. “Immunomodulatory Functions of Human Mesenchymal Stromal Cells Are Enhanced When Cultured on HEP/COL Multilayers Supplemented with Interferon-Gamma.” Materials Today Bio 13 (January 1, 2022): 100194. https://doi.org/10.1016/j.mtbio.2021.100194.https://doi.org/10.1016/j.mtbio.2021.100194http://hdl.handle.net/11603/31934Human mesenchymal stromal cells (hMSCs) are multipotent cells that have been proposed for cell therapies due to their immunosuppressive capacity that can be enhanced in the presence of interferon-gamma (IFN-γ). In this study, multilayers of heparin (HEP) and collagen (COL) (HEP/COL) were used as a bioactive surface to enhance the immunomodulatory activity of hMSCs using soluble IFN-γ. Multilayers were formed, via layer-by-layer assembly, varying the final layer between COL and HEP and supplemented with IFN-γ in the culture medium. We evaluated the viability, adhesion, real-time growth, differentiation, and immunomodulatory activity of hMSCs on (HEP/COL) multilayers. HMSCs viability, adhesion, and growth were superior when cultured on (HEP/COL) multilayers compared to tissue culture plastic. We also confirmed that hMSCs osteogenic and adipogenic differentiation remained unaffected when cultured in (HEP/COL) multilayers in the presence of IFN-γ. We measured the immunomodulatory activity of hMSCs by measuring the level of indoleamine 2,3-dioxygenase (IDO) expression. IDO expression was higher on (HEP/COL) multilayers treated with IFN-γ. Lastly, we evaluated the suppression of peripheral blood mononuclear cell (PBMC) proliferation when co-cultured with hMSCs on (HEP/COL) multilayers with IFN-γ. hMSCs cultured in (HEP/COL) multilayers in the presence of soluble IFN-γ have a greater capacity to suppress PBMC proliferation. Altogether, (HEP/COL) multilayers with IFN-γ in culture medium provides a potent means of enhancing and sustaining immunomodulatory activity to control hMSCs immunomodulation.16 pagesThis item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author.CC BY-NC-ND 4.0 DEED Attribution-NonCommercial-NoDerivs 4.0 Internationalhttps://creativecommons.org/licenses/by-nc-nd/4.0/Text