(188w) Customized, 3D-Printed Devices for Immune Cell Migration across Porous Membranes

Neutrophils are the most abundant immune cell in the body and are the first responders to infections. Important neutrophil functions include chemotaxis, migration and bactericidal function. However, in diabetic patients, these key functions do not occur properly. Diabetics are not only at a higher risk for infection, the severity of the infections are also increased. It is hypothesized that neutrophils in a diabetic environment cannot uptake glucose properly leaving them in an energy deficient state. C-peptide is a 31-chain polypeptide produced in the pancreatic beta cells. Previous work performed in our lab has shown that C-peptide increases glucose absorption into, and ATP release from, red blood cells. Preliminary work has been performed showing that C-peptide also binds to neutrophils in a specific manner. Here, results will show the effect of C-peptide on neutrophil migration. Neutrophils were collected from whole blood samples from consenting adults and separated from other cell types by negative magnetic separation. Neutrophils were incubated with 5 μM cell tracker dye to track migration. The 3D-printed migration device has the same dimensions as a standard 96 well plate with minor adjustments to total height. The transparent device contained eight square spaces for transwell inserts. The transwell inserts will contain a 3-micron membrane to allow neutrophil migration through the pores. One well contained a solution of neutrophils and C-peptide. The opposite well contained 2 μM of fMLP a known chemoattractant, in solution. The migration will be observed with a top down fluorescent microscope and confirmed by flow cytometry.