When analyzing complex biomechanical problems such as predicting the effects of orthopedic surgery, subject-specific musculoskeletal models are essential to achieve reliable predictions. The aim of this paper is to present the Twente Lower Extremity Model 2.0, a new comprehensive dataset of the musculoskeletal geometry of the lower extremity, which is based on medical imaging data and dissection performed on the right lower extremity of a fresh male cadaver. Bone, muscle and subcutaneous fat (including skin) volumes were segmented from computed tomography and magnetic resonance images scans. Inertial parameters were estimated from the image-based segmented volumes. A complete cadaver dissection was performed, in which bony landmarks, attachments sites and lines-of-action of 55 muscle actuators and 12 ligaments, bony wrapping surfaces, and joint geometry were measured. The obtained musculoskeletal geometry dataset was finally implemented in the AnyBody Modeling System™ (AnyBody Technology A/S, Aalborg, Denmark), resulting in a model consisting of 12 segments, 11 joints and 21 degrees of freedom, and including 166 muscle–tendon elements for each leg. The new TLEM 2.0 dataset was purposely built to be easily combined with novel image-based scaling techniques, such as bone surface morphing, muscle volume registration and muscle–tendon path identification, in order to obtain subject-specific musculoskeletal models in a quick and accurate way. The complete dataset, including CT and MRI scans and segmented volume and surfaces, is made available at http://www.utwente.nl/ctw/bw/research/projects/TLEMsafe for the biomechanical community, in order to accelerate the development and adoption of subject-specific models on large scale. TLEM 2.0 is freely shared for non-commercial use only, under acceptance of the TLEMsafe Research License Agreement.