Mesenchymal stem cells (MSCs) have shown broad application prospects in the field of regenerative medicine due to their strong self-renewal ability, low immunogenicity and multi-directional differentiation potential. However, during the in vitro expansion, due to the differences between the culture environment and the physiological state in vivo, MSCs are prone to replicative senescence, which is manifested as slowed proliferation ability, reduced differentiation capacity and weakened function, thereby limiting their clinical transformation and application. Recently, significant efforts have been made to develop effective strategies to delay MSC aging and have made significant progress in gene modification, optimization of culture systems, and application of biomaterials. This review highlights recent advances in approaches to delay senescence in MSCs, aiming to provide a theoretical basis for improving the quality of in vitro culture of MSCs and maintaining their functional characteristics and broaden the scope of MSC applications in tissue engineering and cell-based therapies.