Development of an Alginate Array Platform to Decouple the Effect of Multiparametric Perturbations on Human Pluripotent Stem Cells During Pancreatic Differentiation.

Human embryonic stem cells (hESC)-derived functional cells hold great promise for regenerative cell therapy. Currently approved strategies for clinical translation requires the isolation of the hESCs-derived cells in materials allowing transfer of reagents but preventing integration with the host. However, hESC fate is known to be sensitive to its local microenvironment, both chemical and physical. Given the complexity of hESC response to environmental parameters, it will be important to evaluate the cell response to multiple combinatorial perturbations. Such complex perturbations are best enabled by exploiting high-throughput screening platforms. In this study, the authors report the effect of multivariate perturbations on hESC differentiation, enabled by the development of high throughput 3D alginate array platform. Specifically, the sensitivity of hESC propagation and pancreatic differentiation to substrate properties and cell culture configuration is analyzed. Cellular response to array perturbations is analyzed by quantitative imaging, and cell sensitivity was determined through statistical modeling. The results indicate that configuration is the stronger determinant of hESC proliferation and differentiation, while substrate properties fine-tune the expression around the average levels. This platform allowed for multiparametric perturbations, and in combination with statistical modeling, allows to identify the sensitivity of hESC proliferation and fate to multiparametric modulation.

Temporal control and coordination: the multiple timer model.

We consider the psychological and neurological mechanisms involved in timed behaviors, motor or perceptual tasks that emphasize the temporal relationship between successive events. Two general models for representing temporal information are described. In one model, temporal information is based on the oscillatory activity of an endogenous pacemaker; in the other model, temporal information is interval-based with distinct elements devoted to representing different intervals. We incorporate the interval hypothesis into a process model, the multiple timer model, to account for the timing and coordination of repetitive movements. The model accounts for the patterns of temporal stability observed within each effector and offers a novel account of between-effector coordination. Finally, we consider how timing and temporal coordination may be instantiated in the nervous system.