Stem cells, embryonic and adult, represent the foundation for regenerative medicine. Their unique ability toundergo self-renewal balanced with the capacity to generate numerous differentiated cell types promises torevolutionize therapy of degenerative disorders. A central challenge to their widespread translational use isthe identification of environmental cues and genetic regulatory networks that govern the self-renewal anddirected differentiation of stem cells into a tissue of interest. Identification of the underlying gene regulatorycircuitry at work is a key step for directing stem cell differentiation. Moreover, the ability to modulate andmonitor stem cells to test predicted gene regulatory network relationships is a necessary step in evaluatingplausible, reproducible and efficient methods for the genetic guidance of differentiation.Here we propose to adapt our existing microtechnology platform for dynamic imaging of multiplexedmicroenvironments to understanding and evaluating the role of gene regulatory networks in stem celldifferentiation. The introduction of known genetic activators combined with cell lineage reporters provides aunique readout for the potential use of genetic methods to guide stem cell differentiation. Through the viralexpression of exogenous genetic factors and monitoring of fluorescence reporters we will evaluate generegulatory networks underlying the differentiation of murine embryonic stem cells into pancreatic endoderm,tooth and heart valve tissue as part of the SysCODE consortium. Furthermore, the use of microtechnologytools provide microenvironments that can be exposed to a variety of independent perturbations and theresponse monitored with high temporal and spatial resolution, permitting the evaluation of many geneticinputs along a specific cellular differentiation pathway.The moderate throughput nature of our technology platform in combination with the generation of potentialgene regulatory networks through the SysCODE consortium represent a powerful testbed for the evaluationof network predictions and ultimately potential targets for developing in vitro methods and therapies for tissueregeneration and organogenesis.
|Albrecht, Dirk R; Underhill, Gregory H; Resnikoff, Joshua et al. (2010) Microfluidics-integrated time-lapse imaging for analysis of cellular dynamics. Integr Biol (Camb) 2:278-87|