1. Introduction 1.1 Vascular barriers Functions: Maintain tissue homeostasis, control transport and gas exchange, recruit immune cells during inflammation Figure 1. Schematic illustration of a vasculature barrier in the human body. Endothelial cells as a building element of vasculature barriers line up along the vessel walls and separate the vessel from the tissue. Different stages…
Introduction Here, I will post figure drafts for the paper that introduces the modular µSiM (m-µSiM). Figures Remaining data to obtain: Occludin staining in IMR90-4 from UniBe in µSiM. We have tried this after 2 day culture in µSiM, which worked at UR but is not working at UniBe. Current problem: Occludin is not seen in…
R61; 12 month Progress (Rochester) Small Trans-Agency Blood-Brain Interface Program Investigator Meeting – Agenda_FINAL
Introduction We are differentiating and validating EECM-BMEC (Extended Endothelial Cell Culture Method-Brain Microvascular Endothelial Cell)-like cells in µSiM devices. See this post and this post for method details and validation in traditional cell culturing platforms. Methods Attempt 1 – Overstimulated cells IMR90-4 iPSC EECM-BMEC P3 frozen vial was thawed. At P4 cells were seeded and cultured 2…
Introduction Pages regarding our in situ permeability assay development can be found in these links: Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 1: Approach) Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 2: Experimental Validation) Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 3: Application to Cell…
Competitive ELISA assay to measure the HA shedding from HUVEC monolayer Introduction The endothelial glycocalyx is a dense layer of proteoglycans, glycoproteins, and glycosaminoglycans (GAGs) that helps maintain the integrity of the endothelial cell (EC) barrier.1 The main GAGs are heparan sulfate (HS), chondroitin sulfate, and hyaluronic acid (HA), among which HS and HA are…
Background We at Michigan have been aiming to perform sequential small volume sampling coupled with our ultrasensitive digital immunoassay to provide longitudinal data from the uSIM. We have encountered repeatability problems as well as low molecule abundancy with shortly incubated samples collected from the bottom microfluidic channel. Therefore, COMSOL simulation work has been done to…
Motivation We have discussed the optimization of our in situ permeability assay at length. This assay relies on confocal microscopy, which may not be accessible to all users of the µSiM. Therefore, we developed an alternative permeability assay in the µSiM that is an endpoint, sampling assay. The protocol for this assay can be found…
One of the many contributions Alec S. made to our group is to help us appreciate the importance of the apicobasal (A/B) polarity of endothelial cells. The effects of A/B polarity were demonstrated in his paper: Integrative Biology (2020), 12(11),275–289 where he showed that stimulation of ECs with TNFα from the abluminal chamber caused an…
Introduction As part of our continuing evaluation of the flow cell developed by our collaborators at RIT, our next goal is to demonstrate that neutrophils remain inactivated when flown through a first generation device (Figure 1). For this, we studied neutrophil activation as a function of fluid flow induced shear stress based on the physical…