Controlling neural territory patterning from pluripotency using a systems developmental biology approach.

Successful manufacture of specialized human cells requires process understanding of directed differentiation. Here, we apply high-dimensional Design of Experiments (HD-DoE) methodology to identify critical process parameters (CPPs) that govern neural territory patterning from pluripotency-the first stage toward specification of central nervous system (CNS) cell fates. Using computerized experimental design, 7 developmental signaling pathways were simultaneously perturbed in human pluripotent stem cell culture. Regionally specific genes spanning the anterior-posterior and dorsal-ventral axes of the developing embryo were measured after 3 days and mathematical models describing pathway control were developed using regression analysis. High-dimensional models revealed particular combinations of signaling inputs that induce expression profiles consistent with emerging CNS territories and defined CPPs for anterior and posterior neuroectoderm patterning. The results demonstrate the importance of combinatorial control during neural induction and challenge the use of generic neural induction strategies such as dual-SMAD inhibition, when seeking to specify particular lineages from pluripotency.

Ontogenetic scaling of metabolism, growth, and assimilation: testing metabolic scaling theory with Manduca sexta larvae.

Metabolism, growth, and the assimilation of energy and materials are essential processes that are intricately related and depend heavily on animal size. However, models that relate the ontogenetic scaling of energy assimilation and metabolism to growth rely on assumptions that have yet to be rigorously tested. Based on detailed daily measurements of metabolism, growth, and assimilation in tobacco hornworms, Manduca sexta, we provide a first experimental test of the core assumptions of a metabolic scaling model of ontogenetic growth. Metabolic scaling parameters changed over development, in violation of the model assumptions. At the same time, the scaling of growth rate matches that of metabolic rate, with similar scaling exponents both across and within developmental instars. Rates of assimilation were much higher than expected during the first two instars and did not match the patterns of scaling of growth and metabolism, which suggests high costs of biosynthesis early in development. The rapid increase in size and discrete instars observed in larval insect development provide an ideal system for understanding how patterns of growth and metabolism emerge from fundamental cellular processes and the exchange of materials and energy between an organism and its environment.