In manual assembly systems, workers’ differences in terms of skills, level of expertise and perceived physical effort largely affect the assembly line balancing and system performance. Traditional long-term strategic decisions may not respond to workforce changes and needs, resulting in frequent requests for line rebalancing. In this study, we propose a methodological framework and an easy-to-use Assembly Line Worker Assignment and Rebalancing Problem with different options: workers’ assignment considering their performance variability, integration of worker dependent physical exertion constraints and possibility to use trainers to assist inexperienced workers. A bi-objective linear programming model is proposed aiming to minimise the cycle time and the number of reassigned tasks to respect the initial design while integrating new workers with different characteristics. The -constraint approach is used to build Pareto frontiers for this bi-objective problem. This approach is applied to three real cases. The obtained results show that the developed model can be successfully used in manufacturing companies to help the production managers to deal with workforce turnover and skills heterogeneity.