Date of Award
Master of Science (MS)
Contemporary and traditional methods of perifusional analysis of cells, pancreatic β cells in particular, involve the utilization of costly equipment exhibiting difficulties in setup and inflexibilities in operation and application. Microfluidic devices in general are inexpensive, easy to fabricate and are capable of screening multiple microenvironmental conditions, with the potential to generate concentration gradients for the study of biological phenomena such as cell migration. Utilization of these devices for cellular fluorescence imaging is also possible, paving the way for more sophisticated methods of dynamic assessment. Herein, we fabricated a pre-designed microfluidic device from photolithography, containing microwells capable of immobilizing human β islet or stem cell-derived β cells for perifusional analysis with low (2mM) and high (20mM) concentrations of glucose, following subsequent dynamic estimation of insulin produced using the insulin ELISA. The main aim was to compare and contrast insulin secretory behavior of human islet and stem cell-derived β cells with dynamic study of calcium ion activity during insulin exocytosis through fluorescence imaging, using an alternative method of assessment, namely, microfluidics.
Dennis Barbour Jeffrey Millman Amit Pathak