The CytoR1TM: A Multifunctional Tool for the Separation, Enrichment and Purification of a Targeted Cell Population from a Mixed Sample

R Barua, D Keck, KE Degen, DE Thomas, A Straka, S Manns, K Kinskie, C Coogan, A Nelson, K David, RV Davalos, AR Hyler. The CytoR1: A multifunctional tool for the separation, enrichment, and purification of a targeted cell population from a mixed sample. American Society for Cell Biology Annual Meeting, Washington D.C. December 2022.

The Cyto R1 is an integrated benchtop platform providing multifunctional solutions for sample preparation. The core of the platform is a microfluidic device, Cyto Chip, that utilizes a unique array of over 25,000 insulating microcylindrical posts positioned between two parallel electrodes in order to generate a nonuniform electric field, a technique called dielectrophoresis (DEP). The unique architectural design and positioning of the micro-posts and electrodes facilitates the creation of electrical traps that can be used to capture and hold a targeted cell subpopulation while sustaining a sterile and untouched sample with high viability. This dielectrophoretic Cyto Chip exploits the interaction of the targeted cell with a nonuniform electric field to exploit biophysical properties of cells such as cell size, membrane morphology, and nuclear size. The strength and direction of the DEP force on a targeted cell is dependent on the frequency of the applied electric field as well as the dielectric properties of the cell and suspending media. Preliminary experiments with the Cyto R1 Platform have demonstrated the use of our technology for cell sorting, filtration, and enrichment. The Cyto R1 successfully facilitated the separation of peripheral blood mononuclear cells (PBMCs) from a co-culture with ovarian cancer stem cells (CSCs), both from mice. From a 56.2% CSCs and 43.8% PMBCs co-culture, we were able to deplete 50% of the PMBCs while retaining the original CSC population. Similarly, the Cyto R1 chip has demonstrated its potential for filtering biological debris and satellite glial cells in order to purify a sample of murine trigeminal ganglia (TG) neurons. Lastly preliminary evidence has indicated the use of the Cyto R1 to enrich cell concentrations from 10^3 to 10^5. Ongoing work is the continued development of the Cyto R1’s enrichment capabilities and quantifying the throughput and effectiveness of enrichment.

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