2022 Annual Meeting
Optimizing PLB-985 Migration in Transwell Chemotaxis Assays
This project aims to understand neutrophil migration in the context of infections. Neutrophils are part of the innate immune system and are the most abundant type of white blood cell. They are the first responders to infections and directly interact with pathogens. Though neutrophils are well studied, the key factors that drive neutrophil migration are still unknown. By leveraging CRISPR technology to remove various genes, it is possible to gain more insight into which genes are directly responsible for neutrophil migration. Neutrophils have a short lifespan of 6-10 hours so using CRISPR technology to genetically modify cells would be fairly difficult. A possible solution is to use the PLB-985 cell line (a myeloid cell line) which can be differentiated to behave similarly to neutrophils, and have a much longer lifespan. The goal of this project is to optimize Transwell Chemotaxis Assays in order to study PLB-985 migration with known chemoattractants. Many conditions, such as migration time, pore size, insert type, chemoattractant concentration, and cell concentration, were tested to identify which factors affect migration the greatest. Through various optimization experiments, the conditions that had the most significant effect on migration were found to be migration time, pore size, and chemoattractant concentration.