Heparin/Collagen Surface Coatings Modulate the Growth, Secretome and Morphology of Human Mesenchymal Stromal Cell Response to Interferon‐Gamma

The therapeutic potential of human mesenchymal stromal cells (h‐MSC) is dependent on the viability and secretory capacity of cells both modulated by the culture environment. Our previous studies introduced heparin and collagen I (HEP/COL) alternating stacked layers as a potential substrate to enhance the secretion of immunosuppressive factors of h‐MSCs. Herein, we examined the impact of HEP/COL multilayers on the growth, morphology, and secretome of bone marrow and adipose‐derived h‐MSCs. The physicochemical properties and stability of the HEP/COL coatings were confirmed at 0 and 30 days. Cell growth was examined using cell culture media supplemented with 2 and 10% serum for 5 days. Results showed that HEP/COL multilayers supported h‐MSC growth in 2% serum at levels equivalent to 10% serum. COL and HEP as single component coatings had limited impact on cell growth. Senescent studies performed over three sequential passages showed that HEP/COL multilayers did not impair the replicative capacity of h‐MSCs. Examination of 27 cytokines showed significant enhancements in eight factors, including intracellular indoleamine 2, 3‐dioxygenase, on HEP/COL multilayers when stimulated with interferon‐gamma (IFN‐γ). Image‐based analysis of cell micrographs showed that serum influences h‐MSC morphology; however, HEP‐ended multilayers generated distinct morphological changes in response to IFN‐γ, suggesting an optical detectable assessment of h‐MSCs immunosuppressive potency. This study supports HEP/COL multilayers as a culture substrate for undifferentiated h‐MSCs cultured in reduced serum conditions.