All things cell culture on the μSiM
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…
Introduction In a previous post, an initial version of the flow module and its sealing components were described. In order to improve the initial version based on collaborators’ feedback, several modifications have been made to simplify the device assembly for the end-user (Fig. 1) and address concerns about: 1) Local regions of high shear stress…
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…
Background Previously, we have demonstrated multiplexed digital sensing on MCP1, KC and TNFα secreted by RAW 264.7 cells stimulated in conventional cell culturing well plates. We observed decent TNFα and strong MCP1 response but no KC secretion from RAW cells upon 100ng/ml of LPS stimulation overnight. The work verified our platform’s capability of multiplex cytokine…
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…
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…
Please see the attached .PDF file for a full monocyte isolation protocol. This is for isolating human monocytes from whole blood. Monocyte-Isolation-Protocol
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…
This post is a continuation of project found in Fabricating 0.5 um Channels in Microscope Slides for Staph Invasion Assays. Just to review, after the development of μSiM-CA (Canalicular Array; see Masters et al., Nanomedicine: Nanotechnology, Biology, and Medicine 2019), Jim wanted to develop a platform to answer some more biophysics focused questions. In order…
PDF with all slides: R61 HL154249-01(6 Month Progress)