Increasing label-free stem cell sorting capacity to reach transplantation-scale throughput

Abstract

Dielectrophoresis (DEP) has proven an invaluable tool for the enrichment of populations of stem and progenitor cells owing to its ability to sort cells in a labelfree manner and its biological safety. However, DEP separation devices have suffered from a low throughput preventing researchers from undertaking studies requiring large numbers of cells, such as needed for cell transplantation. We developed a microfluidic device designed for the enrichment of stem and progenitor cell populations that sorts cells at a rate of 150,000 cells/h, corresponding to an improvement in the throughput achieved with our previous device designs by over an order of magnitude. This advancement, coupled with data showing the DEPsorted cells retain their enrichment and differentiation capacity when expanded in culture for periods of up to 2 weeks, provides sufficient throughput and cell numbers to enable a wider variety of experiments with enriched stem and progenitor cell populations. Furthermore, the sorting devices presented here provide ease of setup and operation, a simple fabrication process, and a low associated cost to use that makes them more amenable for use in common biological research laboratories. To our knowledge, this work represents the first to enrich stem cells and expand them in culture to generate transplantation-scale numbers of differentiation-competent cells using DEP. V C 2014 AIP Publishing LLC.

Generation and Analysis of Axiolitic Liquid‐Metal Droplets in a T‐Junction Microfluidic Device

“…In recent years, many methods have been tried, and the majority were simply concentrated on emulsions, encapsulation, sonication, and reactions in solution, which significantly stimulated the advances of the micro‐droplets science and art . Along with the development of microfluidic techniques, droplet‐based microfluidics is becoming a highly useful platform in various areas, such as chemistry, biochemistry, molecular biology, and materials science ,. This system has many promising features, including throughput repeatability, handling small volumes of fluid, low unit cost and so on.…”

3-D In-Bi-Sn Electrodes for Lab-on-PCB Cell Sorting

“…Particles thus experience different dielectrophoretic forces depending on their randomized vertical starting positions in the channel cross section: those nearest the electrodes will experience a strong DEP force, while the rest may slip through the device unsorted by passing through a region of particularly low gradient, e.g., near the channel ceiling, away from the electrodes. Second, many DEP devices with an interdigitated electrode configuration separate cells or particles in batches [4], [7], [8] rather than one by one in a continuous manner, limiting the throughput of such approaches 2156-3950 © 2016 IEEE. Personal use is permitted, but republication/redistribution requires IEEE permission.…”

“…Microfluidic cell and particle separations have been applied to many fields of biological and chemical research including cancer cell isolation for diagnostics [2] and stem cell characterization and enrichment [3], [4]. DEP is arguably among the most mature of these microfluidic label-free cell separation techniques, having been applied to separations involving cancer cells from blood as early as the 1990s [5], [6] and more recently in the enrichment of stem and progenitor cell populations in preparation for transplantation studies [4], [7].…”

“…Finally, both aliquots are recombined at a 1:1 ratio in a lowconductivity DEP buffer consisting of 8.5% sucrose (wt/vol), 0.3% dextrose (wt/vol), and 0.725% Roswell Park Memorial Institute (vol/vol); final conductivity and pH are 100 μS/cm and 7.38, respectively. Low-conductivity buffer is used to enable sorting of biological cells using positive DEP, as shown previously [7], [13], [24]. The final concentration of each cell type is 1E6/ml.…”

Skeletal stem cell isolation: A review on the state-of-the-art microfluidic label-free sorting techniques

“…In their most recent publication (Simon et al, 2014), viable astrocyte progenitor-enriched populations were sorted by DEP at a rate of 2,500 cells per minute. In these devices, interdigitated electrode arrays in a microfluidic chamber were actuated with a 7 V p-p signal at 1 MHz to trap all viable cells.…”