The synthesis of bio polyol from renewable resources has attracted attention in recent years. In particular, it is important to take advantage of bio polyols in the synthesis of polymers. In this study, a series of dimethylformamide (DMF) based polyurethanes were synthesized using polycarbonate polyol/bio polyol (PO3G: polytrimethylene ether glycol prepared from 1, 3-propanediol produced by fermentation from corn sugar), methylene diphenyl diisocyanate (MDI) and 1,4-butandiol (BD). The properties of prepared polyurethane films and the cell structure of wet type artificial leather were investigated. As the bio polyol content increased, the tensile strength of polyurethane films decreased, however, the elongation at break increased significantly. As a result of thermal characteristics analysis, the glass transition temperature of polyurethanes increased when increasing the content of polycarbonate polyol. As a result of comparing the cell characteristics of wet type artificial leathers prepared in this study, it was found that the number and uniformity of cells formed in the artificial leather samples increased when increasing the content of polycarbonate polyol in polycarbonate polyol/bio polyol. From these results, it was found that DMF-based polyurethane containing an appropriate amount of bio polyol could be used for wet type artificial leather. The bio textile analysis system according to ASTM standard was used to measure the bio carbon content of polyurethane. The content of bio carbon increased proportionally with the increase of bio polyol content used in polyurethane synthesis.