Electro-mobility is increasing significantly in the urban public transport and continues to face important challenges. Electric bus fleets require high performance and extended longevity of lithium-ion battery at highly variable temperature and in different operating conditions. On the other hand, bus operators are more concerned about reducing operation and maintenance costs, which affects the battery aging cost and represents a significant economic parameter for the deployment of electric bus fleets. This paper introduces a methodological approach to manage overnight charging of an electric bus fleet. This approach identifies an optimal charging strategy that minimizes the battery aging cost (the cost of replacing the battery spread over the battery lifetime). The optimization constraints are related to the bus operating conditions, the electric vehicle supply equipment, and the power grid. The optimization evaluates the fitness function through the coupled modeling of electro-thermal and aging properties of lithium-ion batteries. Simulation results indicate a significant reduction in the battery capacity loss over 10 years of operation for the optimal charging strategy compared to three typical charging strategies.