Simultaneous cell capture and induction of apoptosis using an anti-CD95 affinity microdevice.
A microfluidic device is designed and demonstrated for the simultaneous capture and induction of apoptosis in Jurkat cells. In this unique case, the cell capture event initiates a biological process. The device features a single channel made from poly(dimethylsiloxane) sealed to a glass substrate. The channel is coated with a series of reagents used in affinity chromatography separations of cells. In this case, the antibody used to capture the cells is functional anti-CD95 which captures the cells by binding to the Fas receptor on the cell membrane and, at the same time, inducing apoptosis via the caspase 8 pathway. Cells retained on the surface of the channel are known to be induced to undergo apoptosis. Medium is flowed slowly through the channel to maintain cell viability while the cells undergo apoptosis. After 3 h, staining with Annexin V-PE and 7-AAD revealed that 43.5% of the cells bound to the anti-CD95 coated channel are apoptotic, whereas 7.9% of cultured Jurkat cells induced with anti-CD95 for 3 h and stained in the same way were determined to be apoptotic by flow cytometry. The device provides a method of determining when apoptosis is induced, maintaining cell viability for long-term analysis and observing cells in real time as they are exposed to reagents that affect apoptosis. In the future, the device will be an invaluable tool for the study of the temporal dynamics of apoptosis.
Reif
Randall D
RD
Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409, USA.
Martinez
Michelle M
MM
Wang
Kelong
K
Pappas
Dimitri
D
D055910:Antibodies, Immobilized / Q000276:immunology*; D017209:Apoptosis; D004867:Equipment Design; D006801:Humans; D019169:Jurkat Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D019014:fas Receptor / Q000276:immunology*
D055910:Antibodies, Immobilized; D019014:fas Receptor