SBIR Phase I: Microfluidic Environment Control for Hepatocyte Bioreactor Optimization

这个小型企业创新研究(SBIR)第一阶段项目将研究一种新型微流控系统在组织培养控制和优化中的应用。原代细胞(来自活的生物体)代表着一个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的微环境要求，目前还没有合适的方法来可靠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势(增加的细胞密度、持续的营养流动、更多的生理传质、更少的细胞/试剂消耗、更高的吞吐能力)是否会为原代细胞培养实验带来更好的平台。本研究将以分离的肝细胞作为主要细胞来源。已知的生化活动(例如葡萄糖消耗和白蛋白合成)将以微流控格式进行测量，并与传统的塑料培养皿进行比较。预计改进的微流控培养条件将导致更高的细胞存活率，更长的培养时间，以及改善肝脏特异性功能。此外，由于该平台的低成本和多元化性质，培养细胞对各种质量传输和可溶性因子条件的响应可以很容易地优化。这项研究的更广泛的影响可以分为四类：基础科学理解、工业药物筛选、组织工程和个性化临床使用。从学术角度来看，提出的原代细胞培养平台可能为细胞生物学的研究提供一种独特的方法。越来越明显的是，细胞的行为不仅仅是由遗传因素决定的，培养环境是细胞信号的主要来源。一个低成本、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。提供更具生理相关性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，需要一种能够充分促进所需功能的组织生物反应器。这里介绍的初步工作将进一步理解微流控技术如何应用于这一领域。最后，这项拟议技术的未来版本可以用于临床应用，这些应用需要对患者的组织进行培养以用于诊断目的。

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.