Characterisation of cellular assemblies in microfluidic systems (synthetic biology to obtain novel antibiotics and optimized production systems)

拟议的研究项目是更大的国际提案SYNMOD（全名：合成生物学以获得新型抗生素和优化的生产系统）的一个组成部分，该项目是与欧洲的五个合作伙伴和英国的一个合作伙伴的合作项目。SYNMOD致力于将合成生物学的基础科学与必要的生物过程工程相结合，以获得新的和改进的抗生素，以克服日益严重的抗菌素耐药性问题。为了实现这一潜力，有必要进行研究，以开发能够快速轻松地培养生物部件和细胞的方案和技术，并了解它们在不同生长条件下的行为。开发这样的技术和协议是这个拟议研究项目的目标。目前，一些工具，如摇瓶，很容易建立一个实验，但提供相对较少的信息，每个实验。其他工具，如具有多个探针的实验室规模的生物反应器，为实验设置成本高且劳动密集型。因此，很难进行充分了解生物部件和细胞行为所需的大量实验。拟议的研究通过使用微制造方法来创建具有透气膜的腔室来提供氧电池所需的解决方案。通过已经制造的用于细胞培养的这种“微流体”反应器系统，它们主要被设计成以“分批”模式操作。这是一种反应器操作模式，其中培养条件在实验期间变化很大。因此，细胞的性质，如大小、组成和功能特性也有很大的不同。相反，在“恒化器”操作模式中，细胞保持在相同的生长条件下，即培养物中的细胞生物量、营养物和产物浓度保持恒定。这是通过受控和连续的营养供应与受控和连续的产物和“过量”细胞的去除相结合来实现的。因此，在“恒化器”操作模式中，将细胞行为与特定生长条件相关联变得可能。在这项研究中，我们建议建立“微流体”反应器系统，在“恒化器”模式下运行。一旦这个问题得到解决，就有可能利用“微流体”系统的一般优势，例如并行化和自动化，来创建一种具有成本效益的工具，可以快速研究和表征细胞行为。

The proposed research project forms an integral part of the larger international proposal SYNMOD (Full name: Synthetic biology to obtain novel antibiotics and optimised production systems), which is a collaborative project with five partners in Europe and one in the UK. SYNMOD seeks to integrate the basic science of Synthetic Biology with the necessary bioprocess engineering in order to achieve new and improved antibiotics to overcome the growing problem of antimicrobial resistance. In order to realise this potential, it is necessary to undertake research to develop protocols and technologies that allow to quickly and easily characterise biological parts and cells, and to understand their behaviour in different growth conditions. Developing such technologies and protocols is the goal of this proposed research project. Currently, some tools, such as shaker flasks, are easy to set up for an experiment, but provide relatively little information per experiment. Other tools, such as bench-scale bioreactors with multiple probes, are costly and labour-intensive to set up for an experiment. It is therefore difficult to perform the large number of experiments required to fully understand the behaviour of biological parts and cells. The proposed research addresses this by using micro-fabrication methods to create chambers with gas permeable membranes to provide the oxygen cells need. Through such 'microfluidic' reactor systems for the cultivation of cells have been fabricated, they have mainly been designed to operate in 'batch' mode. This is a reactor operation mode where the culture conditions vary considerably during an experiment. Accordingly, the properties of the cells, such as size, composition, and functional characteristics vary considerably, too. In contrast, in a 'chemostat' operation mode, the cells are kept in the same growth condition, i.e. the cell biomass, the nutrient and the product concentrations in the culture remain constant. This is achieved by a controlled and continuous supply of nutrient combined with a controlled and continuous removal of product and of 'excess' cells. Therefore, in a 'chemostat' operation mode, it becomes possible to correlate a cellular behaviour with a particular growth condition. In this research, we propose to build 'microfluidic' reactor systems that operate in 'chemostat' mode. Once this is solved, it will then become possible to use the general advantages of 'microfluidic' systems, such as parallelisation and automation, to create a cost-effective tool with which cell behaviour can be rapidly investigated and characterised.