<p>Mesenchymal stem cells (MSCs) playing a crucial role in regenerative medicine due to their multipotent differentiation capabilities and significant paracrine effects. Despite their potential, MSCs face clinical challenges, including low proliferation rates, poor survival post-transplantation, and limited tissue homing. Chitosan, a biopolymer derived from chitin, addresses these challenges effectively due to its biocompatibility, biodegradability, and ability to enhance MSC attachment, proliferation, and survival. Chitosan-based biomaterials, which can be modified through various chemical and physical methods, show substantial promise in regenerative medicine. They can be engineered into forms such as membranes, hydrogels, microgels, scaffolds, nanofibers, and nano- and microparticles and serve multiple applications from three-dimensional in vitro cultures to scaffolds for tissue engineering and in vivo cell delivery systems. Chitosan improves MSC behavior by modulating critical signaling pathways, including Wnt/β-catenin, Notch, and HIF-1α, which are essential for MSC function. Furthermore, adjusting chitosan’s chemical properties can promote specific lineage differentiation and enhance MSC immunomodulatory functions, vital for therapeutic efficacy in inflammatory conditions. Currently, applications of chitosan include wound healing, which will be extended to skin regeneration, bone and cartilage repair, and vascular and neural tissue engineering. Despite progress, challenges in clinical translation persist, particularly concerning safety and standardization. Future research should aim to optimize chitosan biomaterials, refine clinical protocols, and integrate advanced technologies to enhance regenerative outcomes.</p> Graphical Abstract <p>Chitosan-Based Platforms Enhance MSC Viability and Regenerative Function. Chitosan’s degree of deacetylation, molecular weight, solubility, and formulation type modulate their ability to support mesenchymal stem cell (MSC) viability, paracrine activity, and therapeutic performance in regenerative medicine</p> <p></p>