CAN THE OXYGEN TENSION IN A MICROFLUIDIC STEM CELL CULTURE DEVICE BE PRECISELY CONTROLLED DURING BOTH CONTINUOUS AND INTERMITTENT MEDIA EXCHANGE?

使用人类细胞的疗法在解决我们缺乏有效治疗的严重疾病方面有很大的希望。这些疾病特别包括老年慢性退行性疾病，如中风、心力衰竭、老年性失明以及胰岛素依赖型糖尿病和帕金森氏症的晚期。特别是，可以转化为各种特化细胞的干细胞尤其重要。这种转化所需的化学、生物和物理线索是复杂的，需要检查大量的排列才能获得良好的结果。由于所需的许多生化线索的成本非常高，即使是基本的发现研究和培养瓶中的早期翻译研究也经常受到限制，以至于可能错过最佳条件。原则上，可以通过越来越小的烧瓶来解决这个问题，但在实践中，包括去除废养分和添加新鲜养分在内的必要操作变得难以管理。这项拟议的研究通过使用微制造方法创建带有气体渗透膜的腔室来解决这个问题，以提供氧气细胞所需的氧气。我们将使用这种微流控系统，并进一步开发它们，以实现对氧气的最佳控制，这是从干细胞转化为特化细胞的重要线索。该项目将为一项技术奠定基础，最终有可能实现对人类细胞培养最佳条件的更快速和更全面的检查，以生产基础研究和应用研究所需的数量。因为小鼠干细胞的定义特别明确，所以在这项研究中，它们将被用作人类细胞的替代品，以建立原理证明。

Therapies using human cells have great promise for addressing serious medical conditions for which we lack effective treatments. These include particularly those chronic degenerative diseases of old age such as stroke, heart failure, age-related blindness and later stages of both insulin dependent diabetes and Parkinson's disease. In particular, stem cells that can be converted to a variety of specialised cells are especially important. The chemical, biological and physical cues needed for such conversions are complex and it is necessary to examine a large number of permutations to achieve a good outcome. Because many of the biochemical cues required are of very high cost, even basic discovery research and early translational studies in culture flasks are often restricted such that optimal conditions can be missed. In principle it is possible to address this problem by smaller and smaller flasks but in practice the necessary manipulations including removal of spent nutrient and the addition of fresh nutrient becomes difficult to manage. The proposed research addresses this by using micro-fabrication methods to create chambers with gas permeable membranes to provide the oxygen cells need. We will use such microfluidic systems and further develop them to enable best possible control over oxygen which is an important cue for the conversion from stem cells to specialised cells. The project will provide the foundation for a technology with which it will ultimately be possible to achieve more rapid and comprehensive examination of the best conditions for human cell culture to produce the quantities needed for both basic and applied research. Because mouse stem cells are particularly well defined they will be used in this study as a surrogate for human cells to establish the proof of principle.