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Phase Contrast Microscopy Image Correction for Neutrophil Transmigration Studies
Introduction Selective leukocyte interactions with endothelial barriers are a characteristic component of the inflammatory response process. These spatio-temporal interactions can mechanically remodel local tissue, as well as damage healthy tissue with robust inflammation stemming from an over-reactive immune response. This is evident in many pathologies and can complicate severe conditions such as sepsis. Thus, it…
Design and fabrication of the microfluidic flow module
Introduction Sepsis is a serious condition where unregulated systemic inflammation can cause organ failure and death. Recent evidence has also shown that sepsis can cause cognitive impairment in surviving patients. However, the mechanisms that cause sepsis-related damage to the brain are challenging to study with the current in vitro tools. In this project, our goal…
Neural Crest Stem Cell Differentiation into Brain Pericyte-like Cells
Introduction Breakdown of the blood-brain barrier (BBB) is one of the earliest signs of sepsis and is linked with long-term cognitive impairment. As animal models poorly recapitulate human sepsis, alternative approaches are direly needed to understand sepsis-associated brain injury and develop strategies that protect against cognitive decline. To fill this gap, we are developing a…
Collagen IV is deposited below human umbilical vein endothelial cells
Introduction My previous post focused on attempts to reproducibly generate collagen IV with human umbilical vein endothelial cells (HUVECs) so that we would be able to establish protocols for basement membrane protein detection and degradation measurement. Following my presentation on November 18 2020, there were some questions regarding whether or not the observed collagen IV…
Optimizing Cytomix Stimulation on Mono- and Coculture BBB Models
Introduction The µSiM BBB model is being developed at UR with the goal to understand the impact of sepsis on the brain. Here, we use an EECM-BMEC-like cell (iSPC-derived brain endothelial cells) [1] monoculture model and an EECM-BMEC-like cell and brain pericyte-like cell (BPLC; iSPC-derived brain pericytes) [2] coculture model to optimize the concentration of…
Flow Cell/µSiM Testing – Push versus Pull Flow Modes
Introduction Static endothelial cell cultures have been routinely performed and optimized on the µSiM platform for multiple membrane and cell types. In order to increase the physiological relevance of our device with tissue vasculature mimetics, we need to incorporate fluid flow to emulate shear stresses experienced by endothelium under physiological circulatory conditions. Recently Mehran has…