Low-Cost Platform Technology for Rapid Isolation of Peripheral Blood Mononuclear Cells

项目摘要/摘要 CG Science的目标是开发一种低成本的微流控设备，用于快速分离外周血 单个核细胞(PBMC)，基于其专利平台技术，名为“高效” 确定性分离(HEDS)。该第一阶段项目将专注于PBMC，因为它们是必不可少的 细胞用于科学和临床研究的许多领域，包括免疫学、疫苗开发、 药物发现，以及对自身免疫疾病、传染病、移植物抗宿主的研究 疾病和癌症。目前分离PBMC的标准方法是Ficoll密度梯度法 离心法。然而，该方法很难执行，需要大量的实际操作时间，并且 容易出现人为错误和污染。此外，它介绍了偏向和表型的变化 在多次洗涤和离心步骤中，丰富了细胞群并导致细胞损失。这些 缺点使该方法不适合于许多应用，特别是那些需要结果的应用 保持一致、准确并代表疾病状态。提供一种更好的免费方法 从这些缺点，这是具有成本竞争力的菲科尔，CG Science正在开发一种易于使用的 在HEDS技术的支持下，可以在15分钟内从10毫升血液中分离出PBMC的设备。这个 技术具有独特的配置，允许高效的无密度介质隔离细胞和 制造成本低。初步数据显示，HEDS的表现优于Ficoll密度梯度 离心法可显著提高PBMC回收率、RBC去除和血小板耗竭 无法检测到的细胞活力损失--使用简单的重力喂养。这个第一阶段项目的目标是 演示了HEDS的两个最关键的方面--快速、高产量的PBMC分离和低 制造成本-以下3个具体目标：(1)优化HEDS芯片设计，进一步改进电池 恢复和扩展吞吐量，(2)验证优化的芯片性能并表征蜂窝 使用流式细胞术与Ficoll密度梯度离心法进行比较，以及(3)证明 塑料芯片可制造使用软压花纹。研究团队包括发明家 细胞生物学家、塑料设备制造工程师和 经常使用Ficoll的研究和临床实验室。这个项目的成功将导致 开发一种基本工具，有可能取代Ficoll密度梯度作为标准 一种分离PBMCs和许多其他重要细胞类型的方法，并使科学和临床 在许多领域的研究更有效率，更具重复性，更具成本效益。

Project Summary/Abstract CG Scientific aims to develop a low-cost microfluidic device for rapid isolation of peripheral blood mononuclear cells (PBMCs), based on its patented platform technology named “High Efficiency Deterministic Separation (HEDS).” This Phase I project will focus on PBMCs because they are essential cells used in many areas of scientific and clinical research, including immunology, vaccine development, drug discovery, as well as the study of auto-immune disorder, infectious disease, graft-versus-host disease, and cancer. The current standard method for PBMC isolation is Ficoll density gradient centrifugation. However, the method is difficult to perform, requires substantial hands-on time, and is prone to human error and contamination. Further, it introduces biases and phenotypic changes of the enriched cell population and results in cell loss during multiple wash and centrifugation steps. These drawbacks make the method unsuitable for many applications, especially those that require the results be consistent, accurate, and representative of disease states. To provide a better method that is free from these drawbacks and that is cost-competitive with Ficoll, CG Scientific is developing an easy-to-use device that can isolate PBMCs from 10 ml of blood in 15 minutes, enabled by the HEDS technology. The technology has a unique configuration that allows for efficient density-medium-free isolation of cells and low manufacturing cost. Preliminary data have shown that HEDS can outperform Ficoll density gradient centrifugation and deliver significantly better PBMC recovery, RBC removal, and platelet depletion—with undetectable loss of cell viability—using simply gravity feed. This Phase I project is aimed at demonstrating the two most critical aspects of HEDS—rapid, high-yield PBMC isolation and low manufacturing cost—following 3 Specific Aims: (1) optimize HEDS chip design to further improve cell recovery and scale up throughput, (2) validate optimized chip performance and characterize cellular output using flow cytometry in comparison with Ficoll density gradient centrifugation, and (3) demonstrate plastic HEDS chip manufacturability using soft embossing. The investigator team includes the inventor of the HEDS technology, cell biologists, plastic device manufacturing engineers, and directors of research and clinical laboratories that are regular users of Ficoll. The success of this project will lead to the development of an essential tool that will potentially replace Ficoll density gradient as the standard method for isolating PBMCs and many other important cell types, and make scientific and clinical research in many fields more efficient, reproducible, and cost effective.