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

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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.