3D human lung model to study lung disease and formation of fibrosis

用于研究肺部疾病和纤维化形成的 3D 人体肺部模型

基本信息

批准号：
8516142
负责人：
JOAN E NICHOLS
金额：
$ 30.1万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-24 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8516142
关键词：
A549 Absorbable Gelatin Sponge Acute Alveolar Amniotic Fluid Animal Model Apoptosis Biochemical Biological Biological Assay Biology Bioreactors Cell Adhesion Cell Communication Cell Culture Techniques Cell Death Cell Differentiation process Cell Line Cell Proliferation Cell Survival Cell model Cells Collagen Type IV Complex Confocal Microscopy Culture Media Dendritic Cells Dependency Detection Development Disease Elderly Embryo Endothelial Cells Endothelium Environment Enzyme-Linked Immunosorbent Assay Epithelial Cells Epithelium Evaluation Exposure to Fetal Lung Fibroblasts Fibrosis Flow Cytometry Functional disorder Human Human Cell Line Human Engineering Image Imaging Techniques Immune Immune response Immunoprecipitation In Vitro Infection Inflammatory Influenza Influenza A Virus, H1N1 Subtype Influenza A Virus, H5N1 Subtype Injury Label Laboratories Lead Leukocytes Life Lung Lung diseases Measurement Methodology Methods Microscopy Modeling Monitor Morphology Mus Myofibroblast Neuraminidase inhibitor Organ Organ Model Oseltamivir Pathogenesis Pathogenicity Pathway interactions Phenotype Physiology Population Process Production Pulmonary Fibrosis Quantum Dots Rattus Reliance Research Personnel Respiratory physiology Role Serial Passage Somatic Cell Source Stem cells Structure of parenchyma of lung System Technology Testing Tissue Engineering Tissue Model Tissues Toxic effect Toxicology Trachea Umbilical vein Validation Work Wound Healing Xenobiotics alveolar type II cell base cell injury cell motility cell type complex biological systems cytokine drug discovery drug testing efficacy testing embryonic stem cell established cell line human embryonic stem cell human stem cells human tissue immortalized cell improved in vivo lung development lung injury macrophage matrigel microbial monolayer progenitor research study response scaffold stem three-dimensional modeling two-dimensional two-photon whole body imaging

项目摘要

DESCRIPTION (provided by applicant): Current toxicity or pathogenesis studies rely heavily on traditional cell culture methods and animal models. While these systems provide much basic information, a human 3D organ model composed of native cell types interacting in a physiologically relevant way would more closely approximate in vivo conditions. Development of improved in vitro organ models would enable researchers to construct and analyze complex biological systems especially as they relate to organ development, disease pathogenesis, and toxicology and drug discovery. The development of such a model requires identifying the appropriate matrix material, cell types and microenvironment. Our hypothesis is that human embryonic stem or progenitor cells can be cultured on scaffolds to create an in vitro human lung model. It is possible to introduce leukocytes to the system to further mimic the in vivo environment and to study the roles of fibroblasts/myofibroblasts and macrophages/dendritic cells in the development of pulmonary fibrosis. The central hypothesis of this project is that human stem or progenitor cells can be on natural acellular lung or other commercially available scaffolds to create an in vitro human lung model which can be used to study lung disease and the development of fibrosis after microbial infection or exposure to xenobiotic agents. In this project, our aims are to (1) Produce a long term sustainable three dimensional (3D) human pulmonary model and (2) Test the validity of the model using exposure to H5N1, known to cause development of acute conditions and fibrosis in the lung or H1N1 2009 suspected of causing similar but less severe responses. Sham exposure or exposure to circulating strains of influenza A that do not elicit these responses will be used as controls as will addition of neuraminidase inhibitors Tamiflu and Amantidine. In order to produce the best lung model various matrices such as Gelfoam, Matrigel and collagen-IV will be examined for the ability to allow cell adhesion and sustainability. Several matrix materials will be considered, however, acellular (AC) human lung may be the best candidate for a fibrosis model. Recent work in our laboratory has shown this matrix is able to support differentiation of mouse embryonic stem cells toward cells comprising lung tissue with an appropriate morphology. Both AC whole rat trachea-lung and small sections of AC human lung will be tested for efficacy. For a human model, cell sources to be considered include cell lines and primary cells. Among the human cell lines to be trialed are A549 cells (Type II pneumocytes), HUV-EC-C or HUVEC-CS (human umbilical vein endothelial cells developed into a cell line), 13Lu (fetal lung cells) and human embryonic stem cells. Primary and progenitor cells such as HUVEC (primary cells), human amniotic fluid cells, and somatic lung progenitor cells may be used in a supporting role or to populate the matrix. The growth medium will be assessed for efficient proliferation and differentiation of cells into lung tissue.

描述（由申请人提供）：当前的毒性或发病机制研究很大程度上依赖于传统的细胞培养方法和动物模型。虽然这些系统提供了很多基本信息，但由以生理相关方式相互作用的天然细胞类型组成的人体 3D 器官模型将更接近体内条件。改进的体外器官模型的开发将使研究人员能够构建和分析复杂的生物系统，特别是当它们与器官发育、疾病发病机制、毒理学和药物发现相关时。这种模型的开发需要确定适当的基质材料、细胞类型和微环境。我们的假设是，人类胚胎干细胞或祖细胞可以在支架上培养，以创建体外人肺模型。可以将白细胞引入系统中以进一步模拟体内环境并研究成纤维细胞/肌成纤维细胞和巨噬细胞/树突细胞在肺纤维化发展中的作用。该项目的中心假设是，人类干细胞或祖细胞可以在天然无细胞肺或其他市售支架上创建体外人肺模型，可用于研究肺部疾病以及微生物感染或暴露后纤维化的发展外源性制剂。在该项目中，我们的目标是 (1) 制作长期可持续的三维 (3D) 人类肺部模型，以及 (2) 使用 H5N1 病毒暴露测试模型的有效性，已知 H5N1 病毒会导致肺部出现急性病症和纤维化。肺部或 H1N1 2009 疑似引起类似但不太严重的反应。假暴露或暴露于不引发这些反应的甲型流感流行株将用作对照，添加神经氨酸酶抑制剂达菲和金刚烷胺也将用作对照。为了制作最佳的肺模型，将检查明胶海绵、基质胶和 IV 型胶原等各种基质的细胞粘附和可持续性能力。将考虑几种基质材料，然而，无细胞 (AC) 人肺可能是纤维化模型的最佳候选材料。我们实验室最近的工作表明，该基质能够支持小鼠胚胎干细胞向包含具有适当形态的肺组织的细胞分化。 AC 整个大鼠气管肺和 AC 人肺小切片都将进行功效测试。对于人类模型，要考虑的细胞来源包括细胞系和原代细胞。待试验的人类细胞系包括A549细胞（II型肺细胞）、HUV-EC-C或HUVEC-CS（人脐静脉内皮细胞发育成的细胞系）、13Lu（胎儿肺细胞）和人胚胎干细胞。原代细胞和祖细胞例如HUVEC（原代细胞）、人羊水细胞和体细胞肺祖细胞可用于辅助作用或填充基质。将评估生长培养基的细胞有效增殖和分化成肺组织的情况。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Platforms to test the Temporospatial capabilities of carrier Systems in Delivering Growth Factors to benefit vascular bioengineering.

DOI：
10.1016/j.nano.2021.102419
发表时间：
2021-06
期刊：
Nanomedicine : nanotechnology, biology, and medicine
影响因子：
0
作者：
Lissenya B. Argueta;Jean A. Niles;Jason Sakamoto;Xuewu Liu;S. Vega;Luba Frank;Marco Paessler;J. Cor
通讯作者：
Lissenya B. Argueta;Jean A. Niles;Jason Sakamoto;Xuewu Liu;S. Vega;Luba Frank;Marco Paessler;J. Cor

Novel in vitro respiratory models to study lung development, physiology, pathology and toxicology.