Protocol for efficient human MSC chondrogenesis via Wnt antagonism instead of TGF-β

To better implement mesenchymal stem cell (MSC)-based therapy toward cartilage diseases, a more efficient and less off-target chondrogenesis protocol is needed. Here, we present a protocol to induce human MSC chondrogenesis via Wnt antagonism. We describe steps for pellet formation, Wnt antagonism-based chondrogenic induction, and refreshing the differentiation medium. We detail procedures for characterizing MSC chondrogenesis. By using Wnt antagonism instead of conventional transforming growth factor β-based induction, this protocol avoids the potential for induction of chondrocyte hypertrophy/osteogenesis or other lineages. For complete details on the use and execution of this protocol, please refer to Hsieh et al. (2023).¹.