Microfluidic single-cell array platform enabling week-scale clonal expansion under chemical/electrical stimuli

Abstract

Single-cell culture represents the most straightforward method for investigating cellular heterogeneity. In this paper, we present a novel microfluidic platform that can individually array and culture hundreds of cells under chemical and electrical stimuli for week-scale characterization. Single cells can be deterministically and gently captured in a microchamber array on the proposed platform. The size of the microchamber can be adjusted to fit different cell culture times, and this characteristic enables remarkable scalability. Transparent indium tin oxide microelectrodes were integrated with the single-cell array platform for on-chip electrical stimuli. The platform exhibited nearly 90% single-cell efficiency and facilitated week-scale clonal expansion of different types of single cells. Chemical and electrical stimuli affected proliferation and differentiation of MC 3T3-E1 cells were examined on the chip prototype that contained 416 (32 rows × 13 columns) microchambers, and each microchamber had 1 mm diameter. By tracking clonal expansion of cells under chemical/electrical stimuli for relatively long periods, the proposed platform can facilitate the screening of the cell subpopulation with a favorable growth phenotype for drug testing and cell therapy.

Liquid Metal‐Based Multifunctional Micropipette for 4D Single Cell Manipulation

“…The potential applications of the proposed multifunctional micropipettes are broad. For instance, single cell manipulation has been widely used to characterize the molecular genetics of individual cancer cells, tumor cell migration, and characterization of osteogenic differentiation of MC 3T3‐E1 cells . In addition to providing an effective way for single cell trapping and transport, this tool, in particular, allows users to distinguish different types of cells based on their dielectric signatures (based on DEP or Electrorotation responses) without labeling and selectively pick up cells of desired properties .…”

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“…Using this micro sieve, they realized label-free and rapid human breast cancer single-cell isolation with up to 100% trapping yield and >95% sequential isolation efficiency. Luo et al fabricated a high throughput single-cell trap and culture platform to investigate clonal growth of arrayed single-cells under chemical/electrical stimuli for the week-scale period [ 63 ]. To achieve deterministic single-cell capture in large-sized microchambers, a U-shaped sieve with a 5μm-thick bottom microchannel was used to capture a single-cell, and two stream focusing arms were placed in front of the sieve to enhance capture efficiency.…”

Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research

“…Luo et al found that electrical stimulation of pre-osteoblast cells altered cellular differentiation, which was found to be highly heterogeneous. They exposed MC 3T3-E1 (pre-osteoblast) cells to periodical electrical stimulation for short time periods to enhance osteogenic differentiation [ 84 ]. This heterogeneous property of clonal expansion can be used for cell screening and drug testing.…”