An intergrated analytical microfluidic culture system for stem cell research

干细胞提供的潜力，使几乎每一个组织类型的身体在文化中产生和干细胞生物学的主要最新进展引发了新的研究途径，最终将允许临床医生利用这种治疗潜力，以彻底改变再生医学。 然而，目前难以可靠地产生足够数量的这些细胞，主要是因为这些恒定混合的培养群体倾向于在体外以非定向方式分化。 这大大减缓了实现这些细胞的全部临床前景的进展。 最近，已经可以设计微型组织培养“芯片”，其允许数千个单细胞在独立控制的培养基条件下生长并同时分析，实际上允许以单个大规模实验的一小部分成本进行数百个平行实验。 实现这项技术非凡潜力的一个主要挑战是，由于培养体积小，每次实验的细胞很少，因此监测细胞的局部环境以及它们的代谢和分化状态非常有限。 我们建议解决这些挑战的工程一个完全无损的，非接触的，光学分析系统，荧光和拉曼光谱成像的基础上，允许快速，实时，无损，原位分析的细胞和他们的环境。 我们已经组建了一个科学家和工程师团队，他们在干细胞培养、微流体和光谱学等关键技术方面都具有专业知识，以实现和利用这种集成分析微流体培养系统的独特功能。 本文提出的研究的主要目标是详细分析和优化控制单个干细胞分化和扩增的因素，这些因素在当前培养系统中是不可能的。 这项研究将产生关于关键培养基条件和限制这些细胞在培养中扩增和受控分化的其他因素的宝贵信息。 这项工作的结果将导致大规模生产干细胞的创新新方法，并将加速和降低开发重要的新干细胞疗法的成本。

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.