Abstract
SummaryCellular senescence plays an important role in organismal aging and age‐related diseases. However, it is challenging to isolate low numbers of senescent cells from small volumes of biofluids for downstream analysis. Furthermore, there is no technology that could selectively remove senescent cells in a high‐throughput manner. In this work, we developed a novel microfluidic chip platform, termed senescence chip, for ultrahigh‐throughput isolation and removal of senescent cells. The core component of our senescence chip is a slanted and tunable 3D micropillar array with a variety of shutters in the vertical direction for rapid cell sieving, taking advantage of the characteristic cell size increase during cellular senescence. The 3D configuration achieves high throughput, high recovery rate, and device robustness with minimum clogging. We demonstrated proof‐of‐principle applications in isolation and enumeration of senescent mesenchymal stem cells (MSCs) from undiluted human whole blood, and senescent cells from mouse bone marrow after total body irradiation, with the single‐cell resolution. After scale‐up to a multilayer and multichannel structure, our senescence chip achieved ultrahigh‐throughput removal of senescent cells from human whole blood with an efficiency of over 70% at a flow rate of 300 ml/hr. Sensitivity and specificity of our senescence chips could be augmented with implementation of multiscale size separation, and identification of background white blood cells using their cell surface markers such as CD45. With the advantages of high throughput, robustness, and simplicity, our senescence chips may find wide applications and contribute to diagnosis and therapeutic targeting of cellular senescence.
Highlights
Cellular senescence is a state of permanent cell cycle arrest due to genotoxic stresses and has been shown to be involved in organismal aging and tumorigenesis (Campisi & d’Adda di Fagagna, 2007; Munoz-Espin & Serrano, 2014)
We demonstrated the applications of our senescence chips for analysis of senescent cells in human whole blood and mouse bone marrow samples (Figure 4)
After systematically characterizing the performance of our senescence chips with beads and cells, this device was applied to separate spiked human mesenchymal stem cells (MSCs) from whole blood for on-chip cell analysis
Summary
Cellular senescence is a state of permanent cell cycle arrest due to genotoxic stresses and has been shown to be involved in organismal aging and tumorigenesis (Campisi & d’Adda di Fagagna, 2007; Munoz-Espin & Serrano, 2014). Different microfluidic techniques have been developed for cell separation based on their physical properties (size, deformability, density, etc.), including filtration, deterministic lateral displacement (DLD), inertial flow, and acoustofluidics (Chen et al, 2014; Shields, Reyes & Lopez, 2015; Wu, Chen & Lin, 2017; Xavier, Oreffo & Morgan, 2016) Among those techniques, filtration is the most promising approach to process undiluted whole blood for rare cell separation, and scaled up for high throughput (Lin et al, 2010). To overcome the clogging and cell damage issue while still achieve a high throughput and recovery rate, we developed a microdevice (senescence chip) for three-dimensional size sieving by taking advantages of both dead-end flow and cross-flow filtrations. We demonstrated that our scaled-up senescent chip could achieve a parallel processing with a throughput up to 300 ml/hr
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