Sit-to-stand (STS) motion is an important daily activity, and many post-stroke patients have difficulty performing STS motion. Previous studies found that there are four muscle synergies (synchronized muscle activations) in the STS motion of healthy adults. However, for post-stroke patients, it is unclear whether muscle synergies change and which features primarily reflect motor impairment. Here, we use a machine learning method to demonstrate that temporal features in two muscle synergies that contribute to hip rising and balance maintenance motion reflect the motor impairment of post-stroke patients. Analyzing the muscle synergies of age-matched healthy elderly people (n = 12) and post-stroke patients (n = 33), we found that the same four muscle synergies could account for the muscle activity of post-stroke patients. Also, we were able to distinguish post-stroke patients from healthy people on the basis of the temporal features of these muscle synergies. Furthermore, these temporal features were found to correlate with motor impairment of post-stroke patients. We conclude that post-stroke patients can still utilize the same number of muscle synergies as healthy people, but the temporal structure of muscle synergies changes as a result of motor impairment. This could lead to a new rehabilitation strategy for post-stroke patients that focuses on activation timing of muscle synergies.