A Microfluidic-platform Mini-Liver System for Human Liver Biology Studies

用于人类肝脏生物学研究的微流体平台微型肝脏系统

基本信息

批准号：
7918200
负责人：
Michael J Bouchard
金额：
$ 19.05万
依托单位：
DREXEL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-20 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7918200
关键词：
Address Alcohol consumption Alcohols Apoptotic Bile fluid Biological Assay Biological Models Biology Bioreactors Blood Cell Culture System Cell Culture Techniques Cell Line Cells Chemicals Chronic Coculture Techniques Collagen Communication Development Discontinuous Capillary Disease Drug Metabolic Detoxication Endothelial Cells Environment Epidemiologic Studies Ethanol Metabolism Excision Exposure to Extracellular Matrix Future Gallbladder Goals Growth Factor Hepatic Hepatic artery Hepatitis B Hepatitis B Virus Hepatoblastoma Hepatocyte Heterogeneity Homeostasis Human Human Development Infectious Agent Intestines Link Liquid substance Liver Liver diseases Maintenance Malignant neoplasm of liver Metabolism Methods Microfluidics Modeling Molecular Biology Nutrient Organ Oxygen PTK2 gene Pancreas Perfusion Phenotype Phosphotransferases Physiology Portal vein structure Primary carcinoma of the liver cells Proteins Rattus Series Signal Pathway Signal Transduction Simian B disease Simulate Source Spleen Stimulus Study models System Technology Testing Time Toxin Viral Virus Diseases Virus Replication Work alcohol exposure alcohol response bile duct cell type chronic alcohol ingestion liver function novel public health relevance response src-Family Kinases stellate cell

项目摘要

DESCRIPTION (provided by applicant): The liver is the largest organ in the body. It is involved in a myriad of metabolic processes required for body homeostasis and can also detoxify harmful chemicals. The liver receives blood from the hepatic portal vein and the hepatic artery. The hepatic portal vein carries blood from the intestines, pancreas, spleen and gallbladder and exposes the liver to nutrients and toxins derived from these sources. An unfortunate result of the liver's exposure to ingested substances is that some of these substances, such as alcohol, are harmful to the liver. The liver is also a target for persistent viral infections, such as chronic infections with the hepatitis B virus (HBV). Epidemiological studies have linked some types of infectious agents and alcohol consumption with liver diseases and liver cancer; however, attempts to fully characterize many of these diseases in humans have been hampered by the absence of experimentally tractable human liver model systems. Impediments that have limited development of human liver model systems include the difficulty in maintaining differentiated hepatocytes in traditional cell-culture environments and the difficulty in obtaining large numbers of normal human hepatocytes. Hepatocytes are the predominant cell type in the liver and are responsible for many activities that are attributed to the liver; normal hepatocyte function is maintained by communication and contact with other liver cells such as liver sinusoidal endothelial cells (LSEC). The heterogeneity in metabolic processes throughout the liver has generated debate over what defines a functional liver unit. However, even considering this heterogeneity, the liver sinusoid and surrounding cells can be viewed as the most basic structural and functional unit of the liver. This proposal is directed towards: 1.) Developing a novel "mini-liver" culture system using a microfluidic platform that simulates a liver sinusoid functional unit, and 2.) Testing the utility of this system with two focused studies that analyze the effect of hepatitis B virus (HBV) replication and alcohol exposure on specific hepatocyte signaling pathways. The microfluidic platform will have a layered co-culture of hepatocytes and LSECs in a microchannel that mimics a liver sinusoid functional unit. We will initially develop a simple hepatocyte/collagen/LSEC sandwiched configuration and then expand this to include a channel for bile removal and, if necessary, additional sinusoid resident cells such as Kupffer or stellate cells. Exchange of fluid will be regulated through microchannels and external flow control units. We hypothesize that this system will generate an authentic, simple liver sinusoid functional unit. The current proposal focuses on establishing this system and studying the impact of HBV infections and alcohol exposure on hepatocyte physiology. Our long-term goal is to develop a novel human liver model system that can be used to study normal human liver biology. Future use of this system could include studies to understand the impact of other infectious agents on the liver, the consequence of liver exposure to toxins and the effects of various oxygen concentrations on the known heterogeneity of hepatocyte metabolism. PUBLIC HEALTH RELEVANCE: Our understanding of how various human liver diseases develop, such as those associated with persistent viral infections or exposure to toxins or alcohol, has been hampered by the difficulty of acquiring large numbers of human hepatocytes and maintaining their liver-specific functions when grown in traditional cell-culture systems. We propose to use recent advances in bioreactor and microfluidic platform technology to create a novel human liver model system in which small number of human liver cells will be cultured in environmentally controlled microchannels to create a functional liver unit and a novel model for studying human liver functions and diseases.

描述（由申请人提供）：肝脏是人体最大的器官。它参与身体稳态所需的无数代谢过程，还可以解毒有害化学物质。肝脏从肝门静脉和肝动脉接收血液。肝门静脉输送来自肠道、胰腺、脾脏和胆囊的血液，并使肝脏接触来自这些来源的营养物质和毒素。肝脏接触摄入物质的一个不幸结果是，其中一些物质（例如酒精）对肝脏有害。肝脏也是持续性病毒感染的目标，例如乙型肝炎病毒（HBV）的慢性感染。流行病学研究已将某些类型的传染源和饮酒与肝病和肝癌联系起来；然而，由于缺乏实验上易于处理的人类肝脏模型系统，全面表征人类许多此类疾病的尝试受到了阻碍。限制人类肝脏模型系统发展的障碍包括在传统细胞培养环境中难以维持分化的肝细胞以及难以获得大量正常人类肝细胞。肝细胞是肝脏中的主要细胞类型，负责肝脏的许多活动；正常的肝细胞功能是通过与其他肝细胞（例如肝窦内皮细胞（LSEC））的通讯和接触来维持的。整个肝脏代谢过程的异质性引发了关于功能性肝脏单位定义的争论。然而，即使考虑到这种异质性，肝窦和周围细胞仍可被视为肝脏最基本的结构和功能单位。该提案旨在：1.）使用模拟肝窦功能单元的微流体平台开发一种新颖的“迷你肝脏”培养系统，以及2.）通过两项重点研究来测试该系统的实用性，这些研究分析了乙型肝炎病毒（HBV）复制和酒精暴露对特定肝细胞信号通路的影响。微流体平台将在模拟肝窦功能单元的微通道中对肝细胞和 LSEC 进行分层共培养。我们将首先开发一种简单的肝细胞/胶原/LSEC 夹层结构，然后将其扩展为包括用于胆汁去除的通道，并且如果需要，还包括额外的血窦驻留细胞，例如库普弗细胞或星状细胞。流体交换将通过微通道和外部流量控制单元进行调节。我们假设该系统将生成真实、简单的肝窦功能单元。目前的提案重点是建立该系统并研究乙型肝炎病毒感染和酒精暴露对肝细胞生理学的影响。我们的长期目标是开发一种新型的人类肝脏模型系统，可用于研究正常的人类肝脏生物学。该系统的未来使用可能包括研究了解其他传染源对肝脏的影响、肝脏接触毒素的后果以及不同氧浓度对已知的肝细胞代谢异质性的影响。公共卫生相关性：我们对各种人类肝脏疾病如何发展的理解，例如与持续病毒感染或接触毒素或酒精相关的疾病，由于难以获得大量人类肝细胞并维持其肝脏特异性功能而受到阻碍。在传统的细胞培养系统中生长。我们建议利用生物反应器和微流控平台技术的最新进展来创建一种新型的人类肝脏模型系统，其中少量的人类肝细胞将在环境控制的微通道中培养，以创建功能性肝脏单元和研究人类肝功能的新型模型和疾病。