This is the third installment in a series of posts focused on the development of a method for measuring barrier permeability in the µSiM. In Part 1 we discussed the goal of making these measurements in situ at a position beneath the membrane. In Part 2 we validated this approach experimentally on the Andor Dragonfly…
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Introduction This post highlights results from my second attempt to differentiate hiPSCs into endothelial progenitor cells EECM-BMEC-like cells in our lab. For details on methodology, see Nishihara et al., FASEB, 2020 and my previous post. Methods Cells were seeded at 70,000 and 100,000 cells/well on D-3. Only the 100,000 cell/well plate was sorted. Due to media being backordered,…
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Note: a postscript regarding an assay to determine the activity of our MMP-2/MMP-9 inhibitor was added to this post on June 14, 2021. Introduction The blood-brain barrier (BBB) plays an important role in maintaining brain chemistry by restricting the permeability of small molecules and cells. Inflammatory diseases may disrupt the BBB and promote the development…
A quick recap of the goals and initial experiments: https://trace-bmps.org/electrospinning-aligned-polymeric-fibers-on-parylene-stensils/ After obtaining a reproducible optimized fiber deposition settings and an improved device design, we moved on to growing HUVECs on these devices as the next step. As previously stated, given that there is a parallel project with membrane masking of ECM with parylene stencils, we…
In support of the bone infection studies done in the Schwarz lab at URMC, I have recently been tasked with coordinating a new collaboration with Dr. Yates lab in the Chemical Engineering department. The goal of this collaboration is to establish a method to coat our membranes with hydroxyapatite crystals. Two different methods were attempted,…
ALine Capture Device In this post I will go over some of my recent work that is a continuation of Kilean’s research concerning extracellular vesicle (EV) capture and characterization. In the past, this was done using a tangential flow filtration (Figure 1) scheme and clamped devices featuring nanoporous membranes (Figure 2). These devices are straightforward…
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…
Introduction Mural cells, including smooth muscle cells and pericytes, are important for supporting endothelial cells in the vasculature [1]. They play a role in angiogenesis, and contribute to the development of the vascular basement membrane that provides structure for blood vessels [1,2]. Numerous in vitro models of the blood-brain barrier (BBB) have incorporated pericytes alongside…