Theran Suarez, Natalia Alejandra
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Publication Sulfated glycosaminoglycans culture surface for cell manufacture and cell-based applications(2024-04-26) Theran Suarez, Natalia Alejandra; Domenech, Maribella; College of Engineering; Torres Lugo, Madeline; Mora, Camilo A.; Department of Chemical Engineering; Isaza Brando, Clara E.Heparan sulfate (HS) and heparin (HEP) are glycosaminoglycans known for their pivotal roles in regulating and enhancing growth factor availability, influencing various cellular functions including adhesion, proliferation, differentiation, and gene expression. While their individual use in cell culture is recognized for supporting the preservation of both exogenously added and endogenously secreted growth factors, their singular application has limitations. In this study, we propose a novel approach by combining HS and HEP with collagen (COL) to promote cell adhesion through integrin activity. We investigated the effectiveness of rHS/COL at different sulfation levels, demonstrating its capacity to release and regulate Sonic Hedgehog (SHH) ligand concentration by 5.3-fold and enhance ligand signaling strength within 120 hours of cell culture compared to a commercial surface. Additionally, rHS09/COL emerged as a promising alternative substrate for cell culture, ensuring cell growth in serum-free media conditions for 3 to 10 days, surpassing the growth rate of cells cultured on a commercial surface in 2% FBS media. These findings, attributed to the release of rHS/COL ligands and the prevention of growth factor degradation, address the challenges associated with manufacturing mesenchymal stem cells (MSCs), including rapid ligand degradation and the need for constant ligand replenishment to ensure cell growth. Our proposed cell substrate culture offers a cost-effective alternative, overcoming limitations associated with the availability of xeno-free media in the market.