Optimization of muscle cell culturemedia using nonlinear design of experiments

Optimizing media for biological processes, such as those used in tissue engineering and cultivated meat production, is difficult due to the extensive experimentation required, number ofmedia components, nonlinear and interactive responses, and the number of conflicting design objectives. Here we demonstrate the capacity of a nonlinear design-of-experiments (DOE) method to predict optimal media conditions in fewer experiments than a traditional DOE. The approach is based on a hybridization of a coordinate search for local optimization with dynamically adjusted search spaces and a global search method utilizing a truncated genetic algorithm using radial basis functions to store and model prior knowledge. Using this method, we were able to reduce the cost of muscle cell proliferation media while maintaining cell growth 48 h after seeding using 30 common components of typical commercial growth medium in fewer experiments than a traditional DOE (70 vs. 103). While we clearly demonstrated that the experimental optimization algorithm significantly outperforms conventional DOE, due to the choice of a 48 h growth assay weighted by medium cost as an objective function, these findings were limited to performance at a single passage, and did not generalize to growth over multiple passages. This underscores the importance of choosing objective functions that align well with process goals.

Automated summary

The study demonstrates the effectiveness of a nonlinear design-of-experiments (DOE) method in predicting optimal media conditions for biological processes. By reducing the cost of muscle cell proliferation media while maintaining cell growth, the method outperformed traditional DOE, but its applicability is limited to single passages, highlighting the importance of aligning objective functions with process goals.