Engineering zonal human liver functions in vitro using microfluidics

使用微流体在体外工程设计人类区域肝功能

基本信息

批准号：
8773296
负责人：
Salman R Khetani
金额：
$ 8.17万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-22 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8773296
关键词：
Acetaminophen Address Affect Animals Architecture Artificial Liver Biological Biological Assay Cell Culture Techniques Cell Line Cells Chemicals Clinical Coculture Techniques Complex Coupled Cues Data Device Designs Devices Diffusion Disease Drug toxicity Engineering Enzymes Funding Future Gases Generations Germ Cells Grant Hepatic Hepatocyte Heterogeneity Hormonal Hormones Human In Vitro Investigation Knowledge Liver Lobule Long-Term Effects Measures Membrane Microfluidic Microchips Microfluidics Modeling Molecular Outcome Oxygen Pathway interactions Phenotype Physiological Physiology Play Population Heterogeneity Preclinical Drug Evaluation Publications Research Personnel Role Running Stable Populations Staining method Stains Stimulus Stromal Cells System Technology Tissue Model Training Translating Trees Validation design drug metabolism improved in vitro Model in vivo liver function mimicry nitrogen metabolism novel public health relevance response stem cell differentiation three-dimensional modeling validation studies

项目摘要

DESCRIPTION (provided by applicant): Zonal heterogeneity in hepatic functions is well established both in vivo and in vitro, and is an important feature of the liver with implications fr zonal drug toxicity. However, how various factor gradients within the liver interact to modulate zonal hepatic functions in the human liver, and what role liver stromal cells play in such modulation are not well understood. Due to significant species-specific differences in liver functions, inducing and studying factors that regulate zonation in primary human liver cells is critical to elucidate similarities and differences with their animal counterparts for enabling a continuum of in vitro and in vivo investigations. However, in vitro models of human liver zonation are lacking, especially those that evaluate the long-term effects of factor gradients and stromal interactions on zonal functions in primary human hepatocytes. In order to address the aforementioned gap in knowledge, here we propose to investigate the effects of liver- relevant factor gradients and liver stromal cells on long-term functions of primary human hepatocytes in culture. We will utilize microfluidics to enable minimal use of limited primary human liver cells, and create independent, complex and overlapping gradients of soluble gradients onto multicellular liver cultures. We will assess the effects of gradients (O2, hormones) generated by our device on liver functions thought to be zonated in vivo, in the presence or absence of liver stromal cells. Results in vitro will be compared to mechanisms known in vivo. Ultimately, creating approaches and design rules for making a heterogeneous and stable population of human liver cells will allow recapitulation of this important liver feature in systems designed for drug screening, clinical use (i.e. bioartificial liver devices), and stem cell differentiation.

描述（由申请人提供）：肝功能中的区域异质性在体内和体外都得到了很好的确立，并且是肝脏的重要特征，具有带有纬向药物毒性的影响。然而，肝脏中的各种因子梯度如何相互作用以调节人肝脏中的区域肝功能，以及在这种调节中肝脏基质细胞在这种调节中起什么作用。由于肝功能的物种特异性差异，诱导和研究调节原代人肝细胞分区的因素对于阐明与动物对应物的相似性和差异至关重要，以实现体外和体内研究的连续性。然而，缺乏人类肝脏分区的体外模型，尤其是那些评估因子梯度和基质相互作用对原代人肝细胞层次功能的长期影响的模型。为了解决上述知识的差距，我们在这里建议研究肝脏相关因子梯度和肝基质细胞对培养中原代人肝细胞长期功能的影响。我们将利用微流体来最少使用有限的原代人肝细胞，并在多细胞肝培养物上创建独立，复杂和重叠的梯度。我们将评估设备在存在或不存在肝基质细胞的情况下，设备产生的梯度（O2，激素）对被认为是在体内分区的肝功能的影响。将在体外的体外进行比较的结果。最终，创建制造异质和稳定人群人肝细胞种群的方法和设计规则将允许在设计的系统中概括这一重要的肝脏特征药物筛查，临床使用（即生物人工肝脏装置）和干细胞分化。