Monopile-supported offshore wind turbines (OWTs) are dynamically sensitive structures whose fundamental frequencies may be close to those of environmental and turbine-related excitations. The changes in fundamental frequencies caused by pile-soil interaction (PSI) may result in unwanted resonance and serious O&M (Operation and Maintenance) issues, which have been identified as major challenges in the research field. Therefore, a novel model updating framework with an implicit objective function is proposed to monitor both the stiffness and damping variation of the OWT system based on the measured vibration characteristics, which is further verified by laboratory tests. In particular, layered soil was considered in the tests to simulate the practical soil conditions of Chinese seas. Different pile lengths were introduced to consider the long-term PSI effects for rigid piles and slender piles. The results showed that the variation in the fundamental frequency is significantly reduced in layered soil compared with the pure sand scenario. For the OWT systems in layered soil, the variation in foundation stiffness is negatively related to the burial depth under cyclic loading. The proposed model updating framework is proven reliable for support condition monitoring of OWT systems in complicated soil conditions.