Bioprinting as an advanced enabling technology has the capacity to construct tissues with respective anatomical structures. In order to maintain the precise printing resolution for anatomical tissue printing, cell seeding density in bioink is limited. Bone marrow derived mesenchymal stem cells (MSCs) are widely used for cartilage tissue engineering. However, the approach of ideal chondrogenic differentiation of MSCs without hypertrophy still remains elusive. Here, we reported NR2F2 plays a crucial role in MSC chondrogenesis in bioprinted cartilage. NR2F2 over-expressed MSCs showed significantly enhanced chondrogenesis and NR2F2 knockdown cells demonstrated the exactly opposite behavior. We evaluated the cells cultured in monolayer, 3D pellet, and bioprinted 3D scaffold. All observations were consistent among gene expression, biochemical analysis, histological assay, and biomechanical evaluation. The data also revealed possible involvement of NR2F2 in mechanism of MSC chondrogenic differentiation under hypoxic culture condition. Biotechnol. Bioeng. (C) 2016 Wiley Periodicals, Inc.