In this study, Chia seed mucilage (CSM) as a new source and Polyvinly alcohol (PVA) as co-polymer was used to prepare of nanofibers. Different solutions with CSM/PVA volume ratios of 100:0, 80:20, 60:40, 40:60, 20:80, 0:100 were prepared. Results show the increase in the CSM in the polymer solution increased the viscosity, electrical conductivity and viscosity of solution. Scanning electron microscopy (SEM) was to evaluate the morphology of nanofibers. Fourier transform infrared spectrometer (FTIR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA) studies were used to evaluate chemical structure, crystalline structure and thermal characteristics of nanofibers of CSM/PVA. The crystallization index of CSM, PVA, and CSM/PVA nanofibers was 52%, 38%, and 44%, respectively. The non-formation of a new peak and presence of CSM and PVA peaks in the FTIR spectrum of the produced nanofibers are indicative of the lack of chemical reactions between CSM and PVA. Also, the addition of PVA to CSM improves the thermal resistance of the nanofibers.