An integrated microfluidic platform for in situ cellular cytokine secretion immunophenotyping

Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA. .
Lab on a Chip (Impact Factor: 6.12). 08/2012; 12(20):4093-101. DOI: 10.1039/c2lc40619e
Source: PubMed


Rapid, quantitative detection of cell-secreted biomarker proteins with a low sample volume holds great promise to advance cellular immunophenotyping techniques for personalized diagnosis and treatment of infectious diseases. Here we achieved such an assay with the THP-1 human acute moncytic leukemia cell line (a model for human monocyte) using a highly integrated microfluidic platform incorporating a no-wash bead-based chemiluminescence immunodetection scheme. Our microfluidic device allowed us to stimulate cells with lipopolysaccharide (LPS), which is an endotoxin causing septic shock due to severely pronounced immune response of the human body, under a well-controlled on-chip environment. Tumor necrosis factor-alpha (TNF-α) secreted from stimulated THP-1 cells was subsequently measured within the device with no flushing process required. Our study achieved high-sensitivity cellular immunophenotyping with 20-fold fewer cells than current cell-stimulation assay. The total assay time was also 7 times shorter than that of a conventional enzyme-linked immunosorbent assay (ELISA). Our strategy of monitoring immune cell functions in situ using a microfluidic platform could impact future medical treatments of acute infectious diseases and immune disorders by enabling a rapid, sample-efficient cellular immunophenotyping analysis.

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Available from: Weiqiang Chen
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    • "Microfluidics in general provides advantages such as (i) manipulation of liquids and objects at the microscale, (ii) high precision in controlling flow in low Reynolds number regimes (<<1), and (iii) facilitation of high-throughput experimentation by on chip parallelization and greatly reduced volume of expensive reagents and number of cells. Moreover, microfluidic systems have already allowed multiple biological studies including protein crystallization [6], collection of cellular secretions [7], blood circulation [8], angiogenesis [9] and cellular co-cultures [10]. "
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    • "develop microfluidics-based cellular functional immune assays, which is arguably a more direct measurement of the functional status of immune cells. To achieve this, Huang et al. (2012) have recently developed a microfluidic immunophenotyping assay (MIPA) device for rapid and efficient on-chip isolation of peripheral blood mononuclear cells (PBMCs), their stimulation and cellular cytokine secretion measurements (Figure 2B). A key component of the MIPA device is a surface micromachined polydimethylsiloxane (PDMS) microfiltration membrane (PMM) for both isolation of PBMCs from blood and allowing cytokines secreted from lipopolysaccharide (LPS)-stimulated PBMCs to diffuse rapidly into a biosensing chamber for quantitative immunosensing. "
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