SBIR Phase I: Microfluidic Environment Control for Hepatocyte Bioreactor Optimization

这个小企业创新研究（SBIR）第一阶段项目将研究一种新型微流体系统在组织培养控制和优化中的应用。 原代细胞（来自活生物体的细胞）代表了在体外复制生理行为的令人兴奋的机会。然而，由于在生物反应器中维持这些细胞的严格微环境要求，没有合适的方法来可靠地和系统地培养原代细胞。本研究的目的是确定微流控培养平台的优点（增加的细胞密度、连续的营养流、更多的生理质量运输、减少的细胞/试剂消耗、更高的通量能力）是否将导致用于原代细胞培养实验的上级平台。 在本研究中，分离的肝细胞将用作原代细胞来源。已知的生物化学活性（例如葡萄糖消耗和白蛋白合成）将以微流体形式测量，并与传统的塑料培养皿进行比较。预期微流体形式中的改进的培养条件将导致更高的细胞活力、更长的培养时间和改进的肝脏特异性功能。此外，由于该平台的低成本和多功能性，培养细胞对各种质量运输和可溶性因子条件的反应可以很容易地优化。这项研究的更广泛影响可以分为四类：基础科学理解，工业药物筛选，组织工程和个性化临床应用。从学术的角度来看，所提出的原代细胞培养平台可以提供一种独特的方法来研究细胞生物学。 越来越明显的是，细胞行为不仅仅是由遗传因素决定的，培养环境是细胞信号传导的主要来源。低成本、高通量的实验平台将使研究人员能够系统地研究细胞外信号事件对原代细胞行为的影响。从行业的角度来看，目前的药物开发受到快速准确预测药物临床行为的能力的限制。提供生理学上更相关的细胞培养模型的筛选平台将提高选择先导化合物的有效性。随着组织工程的不断发展，有必要拥有一种能够充分促进所需功能的组织生物反应器。 这里提出的初步工作将进一步了解微流体技术如何应用于这一领域。 最后，所提出的技术的未来版本可用于需要培养患者组织以用于诊断目的的临床应用。

This Small Business Innovation Research (SBIR) Phase I project will investigate the application of a novel microfluidic system for tissue culture control and optimization. Primary cells (those derived from living organisms) represent an exciting opportunity to replicate physiologic behaviors in vitro. However, due to the stringent microenvironment demands for maintaining these cells in bioreactors, there is no suitable method to reliably and systematically culture primary cells. The objective of this research is to determine if the advantages of the microfluidic culture platform (increased cell density, continuous nutrient flow, more physiologic mass transport, reduced cell/reagent consumption, higher throughput capability) will result in a superior platform for primary cell culture experimentation. In this research, isolated hepatocytes will be used as the primary cell source. Known biochemical activities (e.g. glucose consumption and albumin synthesis) will be measured in the microfluidic format and compared to traditional plastic dish culture. It is expected that the improved culture conditions in the microfluidic format will lead to higher cell viability, longer culture times, and improved liver-specific functions. Furthermore, due to the low cost and multiplexed nature of the platform, the response of cultured cells to various mass transport and soluble factor conditions can be readily optimized.The broader impacts of this research can be broken down into four categories: basic scientific understanding, industrial drug screening, tissue engineering, and personalized clinical usage. From an academic standpoint, the proposed primary cell culture platform may offer a unique method to study cell biology. It is becoming more and more evident that cell behavior is not determined solely by genetic factors, and that the culture environment is a dominating source of cell signaling. A low cost, high throughput experimental platform will allow researchers to systematically investigate extracellular signaling events on primary cell behavior. From an industry standpoint, current drug development is limited by the ability to rapidly and accurately predict clinical behaviors of drugs. A screening platform that provides a more physiologically relevant cell culture model will improve the effectiveness of selecting lead compounds. As tissue engineering continues to advance, it will become necessary to have a tissue bioreactor that can adequately promote desired functionalities. The initial work presented here will further the understanding of how microfluidic technology can be applied to this field. Finally, a future version of the proposed technology can be used for clinical applications that require the culture of a patient's tissue for diagnostic purposes.