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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…
Monocyte Transmigration in μSiM Devices – Optimizing Microporosity for Future Studies
Introduction Infection or injury of tissue results in inflammatory signalling that motivates leukocyte recruitment to an inflamed location. Monocytes in particular are implicated in both pro inflammatory signalling through the M1 macrophage phenotype and regulation through the M2 macrophage phenotype. The μSiM devices developed in our lab have yet to assess/optimize monocyte transmigration from the…
Human Umbilical Vein Endothelial Cell Shear on Manufactured Hybrid (Dual-Scale) Membranes
ByAlecIntroduction In our labs previously work, difficulties in growing endothelial cells (ECs) under shear flow lead to the exploration of novel membrane technologies that permit firm EC attachment, without compromising membrane permeability or leukocyte transmigration studies. We ultimately landed on the development of effectively ‘dual-scale’ membranes that contained 3 μm pores dispersed amongst an NPN…
Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 1: Approach)
ByJimNOTE: Some of the ideas in this post are borrowed and rephrased from prior posts by Molly and Alec and our paper Khire et al. 2020. The purpose of this thread is to explain things anew with an eye on application in the R61 project. Small molecule diffusion from the well to the channel side…
hiPSC Differentiation into Neural Crest Stem 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…
iPSC-derived Brain Pericyte-like Cells
Introduction Pericytes are mural cells that line the outer surface of microvessels and regulate diverse aspects of vascular development and function. We differentiate brain-pericyte-like cells (BPLCs) from hiPSC clones using the protocol published by Eric Shusta’s lab (Gastfriend et al., 2021) (Figure 1). This protocol describes methods to differentiate hiPSCs to BPLCs via neural crest…