Peanut Shell is a major agro industry waste. Cellulose, the most abundant crystalline component of naturally porous peanut shell biomass. Nanomaterial science has actively used peanut shell as a source of nanocellulose. However, there are few reports on the biocompatibility of cellulose nanocrystal based nanocomposites from agro on mesenchymal cell lines. To evaluate how mesenchymal stem cells behave onto this scaffold in vitro, cellulose nanocrystals (CNCs) made from peanut shells were combined with polyvinyl alcohol (PVA) to create tissue engineering scaffolds. SEM images illustrated that for PVA/CNC nanocomposites and clean PVA respectively, increasing the CNC concentration was accompanied with pore size enlargement. The nanocomposite's three-dimensional porous structure conveyed uneven and intertwined pore structures in additionto the pore distribution. The highest relative crystallinity was observed at 10 wt% of CNCs, according to X-ray diffraction, which also showed that the relative crystallinity of the PVA with 0 and 5 wt% of CNCs was lower than that of neat PVA. In order to confirm the modifications in chemical functional groups, Fourier transform infrared spectroscopy was utilized. The non-cytotoxicity of PVA/CNC_10% was measured for cell viability during an in vitro cytotoxicity test. Additionally, the acquired PVA/biocompatibility CNC's with the Murine Mesenchymal cell line (C 3H 10T 1/2) demonstrated good cell spreading and adherence to the material surfaces. These results suggest that future research into the in vitro integration of mesenchymal cells with a PVA/CNC scaffold can prove to be a promising candidate for regenerative purposes.