Jones, Caroline N.2024-03-142024-03-142022-12December 2December 2https://hdl.handle.net/10735.1/10058With the rise of MRSA (Methicillin-Resistant Staphylococcus aureus), there is a need for investigating alternatives to traditional antibiotics. A growing alternative exists in the form of chemoattraction based therapeutics which enhance the immune system’s response to infection. Novel methodologies will need to be developed to measure the increased activity. Our microfluidic system was used to show that the introduction of chemoattractants increased neutrophil migration towards Staphylococcus aureus, pointing to potential utility in therapy. Traditional migration assays, such as Boyden chambers are not ideal for measuring chemotaxis from mouse cells as they require a large number of cells (>100,000 per condition), make it difficult to pay discern the movements of individual cells, and cannot be used to measure directionality or retrotaxis. Mature neutrophils, band cells, and other cells from the marrow microenvironment were harvested from murine bone marrow and placed in a microfluidic chip in order to measure and quantify their migration patterns. We establish a baseline reaction of migration towards media populated by S. aureus and chemoattractants separately. Then we combined both stimuli to demonstrate how chemoattractants improved immune response and to confirm that this migration rate was four times higher than what neutrophils would respond without the aid of additional stimuli. Experiments found that the average quantity of neutrophils migrating increases when chemoattractants are added to media where S. aureus was present in comparison to media where S. aureus was injected without any chemoattractants. These results confirmed that chemoattractants improved cell migration, offering a potential mechanism for treatment of disease, and showed how our platform was capable of quantifying migrations towards pathogens.application/pdfEnglishEngineering, BiomedicalThesis2024-03-14