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

项目总结/摘要 CG Scientific旨在开发一种用于快速分离外周血的低成本微流体装置 单核细胞（PBMC），基于其名为“高效”的专利平台技术， 确定性分离（HEDS）。第一阶段项目将重点关注PBMC，因为它们是必不可少的 用于许多科学和临床研究领域的细胞，包括免疫学，疫苗开发， 药物发现，以及自身免疫性疾病、感染性疾病、移植物抗宿主 疾病和癌症。目前用于PBMC分离的标准方法是Ficoll密度梯度法 离心法然而，该方法难以执行，需要大量的动手时间，并且 容易出现人为错误和污染。此外，它还介绍了偏见和表型变化的 富集的细胞群并导致在多次洗涤和离心步骤期间的细胞损失。这些 缺点使得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。提供一种更好的方法， 从这些缺点，这是成本竞争力与Ficoll，CG科学正在开发一种易于使用的 该设备可以在15分钟内从10 ml血液中分离出PBMC，这是由HEDS技术实现的。的 该技术具有独特的配置，其允许有效的无密度培养基的细胞分离， 制造成本低。初步数据表明，HEDS可以优于Ficoll密度梯度 离心，并提供显着更好的PBMC回收，RBC去除，和血小板消耗， 无法检测到的细胞活力损失--仅仅使用重力进料。第一阶段项目旨在 证明了HEDS的两个最关键的方面-快速、高产PBMC分离和低 具体目标有3个：（1）优化HEDS芯片设计，进一步提高细胞 恢复和扩大吞吐量，（2）验证优化的芯片性能和表征蜂窝 与Ficoll密度梯度离心相比，使用流式细胞术的输出，以及（3）证明 塑料HEDS芯片的可制造性使用软压花。研究者团队包括发明者 HEDS技术，细胞生物学家，塑料设备制造工程师，以及 研究和临床实验室是Ficoll的常规用户。该项目的成功将导致 开发一种基本工具，有可能取代Ficoll密度梯度作为标准 方法分离PBMC和许多其他重要的细胞类型，并使科学和临床 在许多领域的研究更有效，可重复性和成本效益。

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.