The function of the anterior and posterior cruciate ligaments (ACL and PCL) in the first 120 degrees of flexion has been reported extensively, but little is known of their behavior at higher flexion angles. The aim of this investigation was to study the effects of muscle loads on the in situ forces in both ligaments at high knee flexion (>120 degrees). Eighteen fresh-frozen human knee specimens were tested on a robotic testing system from full extension to 150 degrees of flexion in response to quadriceps (400 N), hamstrings (200 N), and combined quadriceps and hamstrings (400 N/200 N) loads. The in situ forces in the ACL and PCL were measured using the principle of superposition. The force in the ACL peaked at 30 degrees of flexion (71.7 +/- 27.9 N in response to the quadriceps load, 52.3 +/- 24.4 N in response to the combined muscle load, 32.3 +/- 20.9 N in response to the hamstrings load). At 150 degrees, the ACL force was approximately 30 N in response to the quadriceps load and 20 N in response to the combined muscle load and isolated hamstring load. The PCL force peaked at 90 degrees (34.0 +/- 15.3 N in response to the quadriceps load, 88.6 +/- 23.7 N in response to the combined muscle load, 99.8 +/- 24.0 N in response to the hamstrings load) and decreased to around 35 N at 150 degrees in response to each of the loads. These results demonstrate that the ACL and PCL carried significantly less load at high flexion in response to the simulated muscle loads compared to the peak loads they carried in response to the same muscle loads at other flexion angles. The data could provide a reference point for the investigation of non-weight bearing flexion and extension knee exercises in high flexion. Furthermore, these data could be useful in designing total knee implants to achieve high flexion.