An intergrated analytical microfluidic culture system for stem cell research

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Stem cells offer the potential to allow virtually every tissue type in the body to be generated in culture and major recent advances in stem cell biology have sparked new avenues of research that will ultimately allow clinicians to harness this therapeutic potential to revolutionize regenerative medicine. Presently, however, it is difficult to reliably produce these cells in sufficient numbers, principally because these invariably mixed culture populations tend to differentiate in a non-directed manner in vitro. This greatly slows progress toward realizing the full clinical promise of these cells. Very recently, it has become possible to engineer micro-scale tissue culture "chips" that allow thousands of single cells to be grown under independently controlled media conditions and analyzed simultaneously, in effect allowing hundreds of parallel experiments at a fraction of the cost of a single large-scale experiment. A major challenge in realizing the extraordinary potential of this technology is that monitoring the local environments of the cells, as well as their metabolic and differentiation state, is extremely limited due to the small culture volumes and few cells per experiment. We propose to solve these challenges by engineering a completely nondestructive, non-contact, optical analytical system, based on fluorescence and Raman spectroscopic imaging, that permits fast, real-time, nondestructive, in situ analysis of both the cells and their environment. We have assembled a team of scientists and engineers with expertise in each of the key technologies-stem cell culture, microfluidics and spectroscopy-to realize and exploit the unique capabilities of this integrated analytical microfluidic culture system. The primary goal of the research proposed here is to analyze, in fine detail, and optimize the factors that control the differentiation and expansion of single stem cells in ways not possible with current culture systems. This research will yield invaluable information about critical media conditions and other factors that limit the expansion and controlled differentiation of these cells in culture. The results from this work will lead to innovative new approaches for large-scale production of stem cells and will accelerate and reduce the cost of developing important new stem-cell-based therapies.