Dynamic Network Modeling of Stem Cell Metabolism.

Stem cell metabolism is intrinsically tied to stem cell pluripotency and function. Yet, understanding metabolic rewiring in stem cells has been challenging due to the complex and highly interconnected nature of the metabolic network. Genome-scale metabolic network models are increasingly used to holistically model the metabolic behavior of various cells and tissues using transcriptomics data. However, these powerful approaches that model steady-state behavior have limited utility for studying dynamic stem cell state transitions. To address this complexity, we recently developed the dynamic flux activity (DFA) approach; DFA is a genome-scale modeling approach that uses time-course metabolic data to predict metabolic flux rewiring. This protocol outlines the steps for modeling steady-state and dynamic metabolic behavior using transcriptomics and time-course metabolomics data, respectively. Using data from naive and primed pluripotent stem cells, we demonstrate how we can use genome-scale modeling and DFA to comprehensively characterize the metabolic differences between these states.