The objective of the present work was to fabricate poly(vinyl alcohol)-gellan gum nanofiber (PG-NFs) based scaffolds for tissue engineering applications. PG-NFs were fabricated via electrospinning and were characterized using scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR) analysis. Physical properties including water solubility, swelling behavior, contact angle, apparent porosity, biodegradation, and conductivity studies were performed. SEM micrographs displayed long, uniform and randomly oriented PG-NFs of average diameter of 158 ± 23 nm with an interconnected three-dimensional network structure and FTIR study showed gellan gum interaction with PVA through hydrogen bonding. The degradation assay confirmed that as fabricated PG-NFs were stable in the aqueous medium without any significant weight loss. The apparent porosity of PG-NFs was 40%, and conductance was 126.93 pS. The PG-NFs was also proven to be non-toxic and biocompatible by supporting the growth of murine embryonic stem cells (ESCs), similar as control, upon culturing on the same. In summary, stability of PG-NFs in the aqueous medium and significant growth of ESCs in vitro on such 3D nanofibrous scaffolds make it a promising material for various tissue engineering applications. © 2019 Elsevier B.V.