Significant efforts have been applied toward fabricating three-dimensional (3D) scaffolds using 3Dbioprinting tissue engineering techniques. Gelatin has been used in 3D-bioprinting to produce designed 3D scaffolds; however, gelatin has a poor printability and is not useful for fabricating desired 3D scaffolds using 3D-bioprinting. In this study, we fabricated pore size controlled 3D gelatin scaffolds with two step 3D-bioprinting approach: a low-temperature (-10 -°C) freezing step and a crosslinking process. The scaffold was crosslinked with 1-ethyl-3-(3- dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). The pore sizes of the produced 3D gelatin scaffolds were approximately 30% smaller than the sizes of the designed pore sizes. The surface morphologies and pore sizes of the 3D gelatin scaffolds were confirmed and measured using scanning electron microscopy (SEM). Human dermal fibroblasts (HDFs) were cultured on a 3D gelatin scaffold to evaluate the effect of the 3D gelatin scaffold pore size on the cell proliferation. After 14 days of culture, HDFs proliferation throughout the 3D gelatin scaffolds prepared with more than 580 mm pore size was approximately 14% higher than proliferation throughout the 3D gelatin scaffold prepared with a 435 μm pore size. These results suggested that control over the 3D gelatin scaffold pore size is important for tissue engineering scaffolds.