The biologist has been using electrical stimulation via DC, AC, or Hz for cell culture system. However, those electrical stimuli are having inconvenience to use the machine. The enzymatic biofuel cell is one of the alternatives for electrical stimulation in the cell study. This study highlighted that the redox polymer based enzymatic biofuel cell, having suitable concentrations, which provides the biocompatibility and convenient to use for cell culturing. Also, this work suggests what concentration of redox polymer and enzyme should be used to the cellular level. Our enzymatic biofuel cell, including two kinds of traditional osmium redox polymer mediators (PAA-PVI[Os(dmo-bpy)₂Cl] as an anode and PAA-PVI[Os(dCl-bpy)₂Cl] as a cathode), two kinds of enzymes (glucose oxidase (GOx) as an anode and bilirubin oxidase (BOD) as a cathode), and poly(ethylene glycol) diglycidyl ether (PEGDGE) as a cross-linker, were prepared onto 2D screen printed carbon electrode (SPCE). The electrical characterization of the anode and cathode electrodes were examined by cyclic voltammetry (CV) and amperommetry (I-t curve). All electrodes were used after sterilization and we checked the cell viability using CCK-8 and Live&Dead staining. Also, the continuous electrical current under similar condition to the cell culturing were measured by i-t curve. Our enzymatic biofuel cell can be easily immersed into the cell culture media, and it showed good biocompatibility. In this work, we exhibited main consideration when enzymatic biofuel cell applied to the cellular level. The enzymatic biofuel cell will be applied to the in vitro and in vivo system as an electrical stimulating source in the near future.