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Gene expression analysis of HUVECs cultured on uSiM (static vs flow)
Introduction Studies have shown that the shear-stimulation of endothelial cells results in the upregulation of the shear-sensitive transcription factor Kruppel-like factor 2 (KLF2). KLF2 plays a critical role in the regulation of endothelial activity such as the regulation of vascular tone, anti-inflammatory responses, and antithrombotic functions. KLF2 is also responsible for the upregulation of endothelial…
Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 3: Application to Cell Barriers)*
ByJimThis 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…
pedELISA sensing on diffusion limited cytokines in uSIM
IntroductionThe ability to continuously sample and assay for the levels of soluble factors without disrupting the microenvironment is important to acquire sequential secretion profiles in BBB experiments on an uSIM module. Combining small volume sampling, buffer replenishment and sensitive pedELISA technology would enable longitudinal assays of proteins introduced to blood compartment or released from cells….
Monolayer Culture of mouse CMECs on μSIM A-Line Devices
Introduction In the Glading Lab we study a rare disease called Cerebral Cavernous Malformations (CCMs). In this disease, brain capillaries develop greatly enlarged lumens and a disrupted blood-brain barrier. A genetic cause of the disease is the presence of a mutation in KRIT1 that causes loss of KRIT1 protein function in the cells. KRIT1 binds…
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…
Assessing the Permeability of Two Barriers in the Post-capillary Neurovascular Unit Part 2: Measuring R2
ByJimIn Part 1 we explained that the composite barrier in our model of the post-capillary NVU is determined by RBBB = RBlB + R2 + RBrB where is R2 is the resistance of the ‘perivascular space’ in our chip. Unlike the other two, the perivascular space is not a regulated cellular barrier that we need…