A microfabricated platform for cell-based assays: Applications to neural stem cells.

摘要

The development and regulation of tissues and organisms are dependent on the sum of individual cell fate processes. Understanding the mechanisms by which external and intrinsic cues lead to distinct cell fates such as proliferation, differentiation, and apoptosis will be crucial to the success of cell-based therapies in the treatment of human disease. Currently, the tools available to study the role of the cellular microenvironment on cell fate processes are relatively limited---in part, due to the number of interdependent variables (soluble factors, cell-cell interactions, cell-matrix interactions). Investigation of these variables using conventional cell-based assays can be prohibitive in terms of cost and/or time. Automation of cell-based assays has already begun to be used in the industrial realm; however, these commercial instruments are beyond the means and needs of most basic research labs. Microfabricated platforms for cell-based assays may provide a simple and economical alternative for conducting parallel studies of cell fate at the bench-scale.; In this work, we have developed and characterized a microfabricated platform, applied it to that is amenable to the study of neural progenitors. First, a robust and versatile cell culture platform employing a three-dimensional SU-8 microfabricated array was designed, modeled, and built. Next, the microfabricated array monitored the proliferation and differentiation of approximately 1000 neural progenitor cells per array in response to soluble growth factors and cell-cell interactions. 3 prototypical mitogens were shown to affect the proliferative behavior of neural progenitors in distinct patterns. In the course of this monitoring, we also identified a rare cell phenotype in the neural progenitor cultures. We characterized similar cells, termed adult hippocampal progenitor associated cells, to determine their identity and ability to give rise to progenitors. Collectively, these studies established a novel application of micro fabrication to cell-based assays, provided insight into the role of the mitogens on the proliferative behavior of neural progenitors, and detected the low-frequency generation of neural progenitor-like cells from a novel cell source. The application of our high content cell-based assays, integrated with traditional biological approaches, illustrates the potential of this platform to detect subtle changes in heterogeneous cell behavior.