VEC3-Valve Enabled Cell Co-Culture Platforms for Cancer Biology Study

用于癌症生物学研究的 VEC3-Valve 支持细胞共培养平台

基本信息

批准号：
8153605
负责人：
Jin Chen
金额：
$ 16.16万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-12 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8153605
关键词：
Address Affect Behavior Biochemical Biological Assay Biological Process Blood Vessels Cancer Biology Cell Communication Cell Culture System Cell Culture Techniques Cell Density Cells Coculture Techniques Communication Culture Media Culture Techniques Data Development Device Designs Devices Disease Endothelial Cells Engineering Environment Ephrin-A1 Event Extravasation Fibroblasts Glucose Goals Guanine Nucleotide Exchange Factors Hippocampus (Brain)Histologic Hypoxia Immigration Incubators Individual Lead Life Ligands Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Methods Microfluidic Microchips Microfluidics Microscope Microscopy Molecular Neoplasm Metastasis Neurons Nutrient Oncogene Activation Output Oxygen Performance Physiological Play Population Process Protocols documentation Reporting Research Research Project Grants Role Signal Transduction Solutions Staining method Stains Targeted Research Technology Time Tumor Angiogenesis Tumor Suppressor Proteins Vascular Endothelial Growth Factors anticancer research base cancer therapy cell behavior cell motility cell type cellular imaging design improved in vivo interest migration neoplastic cell novel novel therapeutic intervention operation programs receptor response success synaptogenesis time use tumor tumor progression user-friendly

项目摘要

DESCRIPTION (provided by applicant): It is becoming increasingly clear that the tumor microenvironment plays a key role in tumor progression, pointing to a need to develop technologies to study tumor cell behavior in different microenvironments. Toward this goal, we have created Valve Enabled Cell Co-Culture (VEC3) platforms, which are a new class of microfluidic devices designed for analyzing interactions between tumor cells and others in the tumor microenvironment. The technology enables separate culture of distinct cell types and cell-cell interactions through either soluble factors or physical contacts between spatially separated cell populations while maintaining fluidic control over their individual culture environment. In addition, through selective blockage of the exchange of specific ligands between distinct cell populations, the platform can be used to identify the functions of relevant ligands of interest. Traditional cell co-culture techniques and reported microfluidic cell co-culture platforms have limitations and cannot address all important cell co-culture needs. The proposed VEC3 cell co-culture platform, through the introduction of a simple, robust, and user-friendly pneumatically or hydraulically controlled valve to reversibly separate or connect adjacent cell culture chambers, not only allows for separate culture and treatment of individual cell types, but also permits real-time, live-cell imaging of cellular interactions. To date, as a proof of principle, VEC3 has been applied to observe dynamically synapse formation between hippocampal neurons, analyze tumor-endothelial interactions in normoxic and hypoxic environments, study tumor-fibroblasts interactions in 3D matrices, and quantify tumor-endothelial cross migration mediated by various molecules. In the proposed research we will further develop VEC3 through quantitative characterizations of cellular microenvironments, including cell density and uniformity, glucose and oxygen concentration, and cell-cell interaction rates. Through improved design and performance engineering, we will develop optimized platforms and operation protocols to increase the success rate of VEC3-based assays. In addition, we will implement new functions such as controlled cell-cell interactions via blockage of the exchange of specific ligands between two cell populations. More importantly, we will apply VEC3 to study tumor- endothelial cross migration mediated by various ligands and receptors and identify the functions of specific ligands in cell-cell interactions. Therefore, successful execution of the proposed research will lead to a new class of versatile, multifunctional VEC3 microfluidic platforms that are widely applicable to cancer biology. This device will be used to elucidate the molecular mechanisms underlying tumor angiogenesis, intravasation and metastasis, which could eventually lead to better cancer treatments. The overall quantitative milestone is to achieve 95% success rate in assays using the VEC3 platforms for tumor angiogenesis, intravsation and metastasis studies with quantitative parameters of cellular communication. PUBLIC HEALTH RELEVANCE: Cell migration is critical for many biological processes, and tumor-endothelial cross-migration is of fundamental importance to tumor angiogenesis. The goal of this research project is to develop novel microfluidic cell co- culture platforms that allow for the quantitative assessment and identification of the molecular mechanisms that regulate cell migration and tumor-endothelial interactions. This will lead to new therapeutic approaches for treating various diseases, such as cancer, that arise from aberrant cell migration.

描述（由申请人提供）：越来越清楚的是，肿瘤微环境在肿瘤进展中起着关键作用，这表明需要开发技术来研究不同微环境中肿瘤细胞的行为。为了实现这一目标，我们创建了 Valve Enabled Cell Co-Culture (VEC3) 平台，这是一类新型微流体设备，旨在分析肿瘤细胞与肿瘤微环境中其他细胞之间的相互作用。该技术能够通过可溶性因子或空间分离的细胞群之间的物理接触实现不同细胞类型的分离培养和细胞间相互作用，同时保持对其个体培养环境的流体控制。此外，通过选择性阻断不同细胞群之间特定配体的交换，该平台可用于识别相关感兴趣配体的功能。传统的细胞共培养技术和报道的微流控细胞共培养平台具有局限性，不能满足所有重要的细胞共培养需求。所提出的VEC3细胞共培养平台，通过引入简单、坚固且用户友好的气动或液压控制阀来可逆地分离或连接相邻的细胞培养室，不仅允许单独培养和处理单个细胞类型，而且还可以对细胞相互作用进行实时活细胞成像。迄今为止，作为原理证明，VEC3 已应用于动态观察海马神经元之间的突触形成，分析常氧和缺氧环境中的肿瘤 - 内皮相互作用，研究 3D 矩阵中的肿瘤 - 成纤维细胞相互作用，并量化肿瘤 - 内皮细胞交叉迁移介导的相互作用由各种分子。在拟议的研究中，我们将通过细胞微环境的定量表征进一步开发 VEC3，包括细胞密度和均匀性、葡萄糖和氧气浓度以及细胞间相互作用率。通过改进设计和性能工程，我们将开发优化的平台和操作协议，以提高基于 VEC3 的检测的成功率。此外，我们将实现新功能，例如通过阻断两个细胞群之间特定配体的交换来控制细胞间相互作用。更重要的是，我们将应用VEC3来研究各种配体和受体介导的肿瘤-内皮交叉迁移，并鉴定特定配体在细胞与细胞相互作用中的功能。因此，成功执行拟议研究将产生一类新型多功能、多功能 VEC3 微流体平台，广泛适用于癌症生物学。该装置将用于阐明肿瘤血管生成、内渗和转移的分子机制，最终可能带来更好的癌症治疗。总体定量里程碑是使用 VEC3 平台通过细胞通讯定量参数进行肿瘤血管生成、血管内血管生成和转移研究的检测成功率达到 95%。公共卫生相关性：细胞迁移对于许多生物过程至关重要，肿瘤内皮交叉迁移对于肿瘤血管生成至关重要。该研究项目的目标是开发新型微流体细胞共培养平台，以定量评估和识别调节细胞迁移和肿瘤内皮相互作用的分子机制。这将带来新的治疗方法来治疗由异常细胞迁移引起的各种疾病，例如癌症。