Many biological systems, such as cell cultures in vitro, are complex, non-linear, and time varying, resulting in challenges for the development of effective control methods. Therefore, it is beneficial to develop simulation environments to understand dynamics of these biological systems and, for example, compare different control strategies. In this paper, we created a mathematical model to simulate temperature-dependency of beating human cardiomyocyte cultures. The developed system can be used for comparing different closed-loop control strategies and for studying how sensitive the beating of cardiomyocytes is for temperature variations. We employed the system for comparing the performance of fuzzy logic-based and traditional Proportional-Integral controllers and presented, how their performances varied in different cell culture cases. Our results indicate clearly, how the fuzzy logic-based controller can outperform the traditional Proportional-Integral controller when they are implemented to control the beating of cardiomyocyte cultures.Note to Practitioners-This paper was motivated by the challenge of creating controllable engineered living materials. For this reason, we created a simulation environment to help the development of these materials, thus to reduce the discovery time. As a demonstration, we showed in this paper how the developed simulation environment can be used to design and compare different controller strategies, such as the traditional Proportional-Integral-Derivate controller and controllers based on fuzzy-logic. In the paper, we implemented different controllers and cardiomyocyte environments to study how beating of cardiomyocyte cultures could be controlled in an optimal way in vitro. We show, how fuzzy-logic controllers outperform the traditional PI-controller when applied to a highly non-linear and time-varying biological system.