Spheroid aspiration-assisted 3D bioprinting is a promising technique for creating tissue spheroids in vitro, aiding tissue engineering and regenerative medicine. This project examines the growth of bioprinted spheroids from 2D to 3D cultures. Our findings show successful 3D-printed spheroids with accurate structural and functional characteristics. This advancement enhances disease modeling, drug screening, and regenerative therapies, paving the way for breakthroughs in personalized medicine and improved medical research.