Integrated Microfluidic Device for Functional Secretory Immunophenotyping of Immune Cells

Abstract

Integrated platforms for automatic assessment of cellular functional secretory immunophenotyping could have a widespread use in the diagnosis, realtime monitoring, and therapy evaluation of several pathologies. We present a microfluidic platform with integrated biosensors and culture chambers to measure cytokine secretion from a consistent and uniform number of immune cells. The biosensor relies on a fluorescence sandwich immunoassay enabled by the mechanically induced trapping of molecular interactions method. The platform contains 32 cell culture chambers, each patterned with an array of 492 microwells, to capture and analyze both adherent and nonadherent immune cells. Multiple stimuli can be delivered to a set of culture chambers. Per chamber, we were able to capture consistently 1113 ± 191 of blood-derived monocytes and neutrophils and 348 ± 37 THP-1 monocytes. Good occupancy efficiencies of ∼70% with a uniformity of ∼90% across all of the culture chambers of the device were achieved. Furthermore, we demonstrate that up to 96% of cells remain viable for the first 48 h. The employment of epoxy-modified glass substrates and active mixing enhanced the biosensing performance compared to the use of bare glass and simple diffusion. Finally, we performed functional secretory analysis of interleukin-8 and tumor necrosis factor alpha from human neutrophils and monocytes, stimulated with various doses of lipopolysaccharide and phorbol 12-myristate 13-acetate−ionomycin, respectively. We foresee the employment of our microfluidic platform in the diagnosis of different pathologies where alterations in cytokine secretion patterns can be used as biomarkers.

Micro/Nanofluidic‐Enabled Biomedical Devices: Integration of Structural Design and Manufacturing

“…For another example, tumor necrosis factor alpha (TNFα) is an important biomarker in type II diabetes, [ 129 ] and its detection is the focal point of the microfluidic device. [ 130 ] Figure 9C shows an electrochemical biomarker detection device for TNFα using a sandwich immunoassay. [ 131 ] The device uses polymeric fluidics for sample control, inlet and outlet syringe ports, and two working electrodes for electrochemical signal and qualitative testing with antibodies.…”

Pancreas-on-a-Chip Technology for Transplantation Applications

“…Immunosuppressive medication which is an inevitable part of the islet transplantation could also get benefit from OoC technologies. OoC devices can provide a platform for testing the compatibility of the immunosuppressive regimen with immune system [66][67][68][69], as well as the effect of the immunosuppressive reagents on isolated islet function and viability [70]. OoC technologies have been used for studying the effect of immunosuppressive medications, tacrolimus, and cyclosporine A on T cell interactions and adhesion to endothelial adhesion molecules [71].…”

“…OoC technologies have been used for studying the effect of immunosuppressive medications, tacrolimus, and cyclosporine A on T cell interactions and adhesion to endothelial adhesion molecules [71]. Rodrigues-Moncayo et al developed a microfluidic chip with integrated biosensors that automatically measures interleukin-8 and tumor necrosis factor-alpha secretion from a consistent number of blood-derived monocytes, neutrophils, and THP-1 monocytes [66]. Such devices could be used for studying instant blood-mediated inflammatory reaction (IBMIR) and auto-and allo-immunity.…”

Ultrasensitive Multiparameter Phenotyping of Rare Cells Using an Integrated Digital‐Molecular‐Counting Microfluidic Well Plate

“…[ 30 ] The difficulty of assaying cell secretion ex vivo is compounded by the need to study the responses of small number of cells to multiple conditions, further dividing the number of cells available per assay if robust and reproducible conclusions are to be reached. Integrated microfluidic cellular secretory platforms monitoring collective cell cytokine secretion response have been widely explored to realize higher measurement throughput, [ 31,32 ] lower cell and reagent usage per measurement [ 31–40 ] and increased assay speed. [ 34–37,40 ] These microfluidic platforms incorporate microscale immunosensors based on localized surface plasmon resonance, [ 35,36,40 ] mechanically induced trapping of molecular interactions, [ 32 ] bead‐based amplified luminescent proximity homogeneous assay [ 34,37 ] or antibody arrays.…”

“…Integrated microfluidic cellular secretory platforms monitoring collective cell cytokine secretion response have been widely explored to realize higher measurement throughput, [ 31,32 ] lower cell and reagent usage per measurement [ 31–40 ] and increased assay speed. [ 34–37,40 ] These microfluidic platforms incorporate microscale immunosensors based on localized surface plasmon resonance, [ 35,36,40 ] mechanically induced trapping of molecular interactions, [ 32 ] bead‐based amplified luminescent proximity homogeneous assay [ 34,37 ] or antibody arrays. [ 31,33,38,39 ] Despite their integrability in a microfluidic platform, these immunosensors still suffer from limited sensitivity which still prevents the microfluidic analysis from tackling the challenges in rare cell clinical studies as presented above.…”