Monocyte Isolation via CD14+ Selection (Protocol)
Please see the attached .PDF file for a full monocyte isolation protocol. This is for isolating human monocytes from whole blood. Monocyte-Isolation-Protocol
-
-
hiPSC Differentiation to Endothelial Cell Culture Method, Brain Microvascular Endothelial Cells (EECM-BMEC-like cells), Round 1
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…
-
R61: Slide Decks for 6 Month Progress Report
PDF with all slides: R61 HL154249-01(6 Month Progress)
-
Collagen IV deposition by hCMEC/D3 (endothelial cell) and HBVP (pericyte) co-cultures in the μSiM-hNVU
Note: A postscript regarding a gelatin assay to determine MMP-9 activity was added to this post on Feb 25, 2021. Introduction The blood-brain barrier (BBB) plays an important role in maintaining brain chemistry by restricting the permeability of undesirable small molecules and cells. Inflammatory diseases may disrupt the BBB and promote the development of neurological…
-
Introduction of beads into the flow module
1. Introduction Fluid flow is one of the key elements of vascular models for delivering nutrients and growth factors to cells as well as introducing other components like neutrophils. In post 1, we described our modular design for incorporating a flow module into the uSiM device. Then, we presented fluid flow simulation and its experimental…
-
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…
-
Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 2: Experimental Validation)
This is the second 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. Because our preference is to use the trench down configuration in the…
-
hiPSC Differentiation into Brain Pericyte-like Cells, Round 2
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…
-
HBVP (pericyte) and hCMEC/D3 (endothelial cell) µSiM Coculture Optimization
Introduction Breakdown of the blood-brain barrier (BBB) is one of the earliest signs of sepsis and is linked with long-term cognitive impairment. We are developing a coculture model of the human blood-brain barrier (µSiM-hBBB) to elucidate mechanisms of BBB breakdown during sepsis. The first version of the µSiM-hBBB will be composed of two cellular components…
-
iPSC IMR90-4 ApoE Genotyping
Introduction The ApoE4 allele of the apolipoprotein E (ApoE) gene is the greatest known genetic risk factor for Alzheimer’s disease. Further it has been linked to increased susceptibility and duration of infection. On the other hand, the ApoE3 allele has been shown to have a protective effect against severe sepsis. Importantly, ApoE has an important role…
