Resumen: Mesenchymal stem cells (MSC) are recognized for their immunomodulatory effects and regenerative properties, being promising therapeutic agents for a wide range of diseases. To ensure a localized effect of MSC in the organism biobased hydrogels have been tested for their ability to act as a matrix-embedded to improve MSC targeted delivery. In this context, nanocellulose (NC) has been used for drug delivery, showing biocompatibility and durability in time, but until now NC has not been tested for MSC growth exploiting the size and aldehyde content of NC. In this study, cellulose nanocrystals (CNC), cellulose nanofibers (CNF) and microfibrillated cellulose (MFC) were studied after one-pot oxidation and further crosslinking with chitosan (mass ratio 1:5). Size and aldehyde content of oxidized NC samples were evaluated to analyze their influence on the hydrogel’s properties. The crosslinked hydrogels were analyzed by FESEM, swelling ability, FTIR, compression tests, thermal stability, and stability in culture cell conditions. Oxidized-MFC hydrogel improved the mechanical stability and swelling behavior, but it lacks stability at cell conditions possibly due to its low aldehyde content (0.54 mmol/g). Conversely, oxidized CNF and oxidized CNC formed suitable crosslinked hydrogels for cell adhesion, and for growing and proliferating of MSC 3D spheroids after 120 h. However, only hydrogel with PO-CNF/chitosan shows antibacterial activity as well as MSC proliferation. Graphical abstract: [Figure not available: see fulltext.]