Regenerated cellulose (RC) bio-nanocomposite fibers reinforced with halloysite nanotubes (HNT) were fabricated through wet spinning technique via ionic liquid as a green solvent. Mechanical properties, water uptake, thermal stability, and cytocompatibility of the obtained fibers were examined. FTIR spectra indicated the uniform dispersion of HNT in the cellulose network. XRD analysis, together with FE-SEM images indicated that HNT was dispersed homogenously in the polymer. Moreover, mechanical and thermal stabilities of the nanocomposite fibers were notably increased through the addition of HNT. Eventually, human skin fibroblasts proliferation on nanocomposite fibers demonstrated good cyto-compatibility. These findings highlight the potential of HNT nanocomposite fibers for biological and biomedical applications.