All things cell culture on the μSiM
Introduction We are investigating some issues seen with long term attachment (up to 6 days) of the EECM-BMEC-like cells seen by collaborators in Bern, where they noticed black aggregates on the coated control devices that get worse at day 6. Some attachment issues and aggregates have also been seen at the University of Rochester in…
Overview Recently, we performed a series of flow cytometry studies assessing neutrophil (PMN) activation in first generation flow devices. All the relevant data and can be found in a previous post: here. As part of the analysis, we chose to quantify relative PMN activation by comparing isotype control histograms against L-selectin (CD62L) expression histograms. This…
Introduction Vascular hyperpermeability and hemorrhagic stroke are hallmark symptoms of Cerebral Amyloid Angiopathy (CAA), a cerebrovascular disease characterized by amyloid-β deposition around the brain microvasculature. In CAA pathogenesis, vascular Aβ accumulation contributes to structural alterations and loss of integrity in the neurovascular unit/blood-brain barrier (NVU/BBB) resulting in “leakier” blood vessels, intracerebral hemorrhages, increased risk of…
Slide deck from Large Group Meeting 12-21-12 (PPT) Access requirements for the AFM cantilver to contact the cells requires the cells to be grown on the membrane surface in what is typically the bottom channel of A-line devices (Typical A-line configuration is shown in slide 2). A PDMS block is attached to the well side…
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 of neurological disorders [1]. One potential mechanism of BBB disruption is the degradation of the extracellular matrix proteins that make up the endothelial…
Background From previous experimental results as well as COMSOL simulations presented in our older posts, we observed a universal non-uniform analyte distribution in the μSIM bottom channel due to limited lateral diffusion rate. This uneven distribution profile will limit our sensitivity in using small volumes of supernatants collected from the ports which are far away…
1. Introduction 1.1 Vascular barriers Functions: Maintain tissue homeostasis, control transport and gas exchange, recruit immune cells during inflammation Figure 1. Schematic illustration of a vasculature barrier in the human body. Endothelial cells as a building element of vasculature barriers line up along the vessel walls and separate the vessel from the tissue. Different stages…
Introduction Here, I will post figure drafts for the paper that introduces the modular µSiM (m-µSiM). Figures Remaining data to obtain: Occludin staining in IMR90-4 from UniBe in µSiM. We have tried this after 2 day culture in µSiM, which worked at UR but is not working at UniBe. Current problem: Occludin is not seen in…
R61; 12 month Progress (Rochester) Small Trans-Agency Blood-Brain Interface Program Investigator Meeting – Agenda_FINAL
Introduction We are differentiating and validating EECM-BMEC (Extended Endothelial Cell Culture Method-Brain Microvascular Endothelial Cell)-like cells in µSiM devices. See this post and this post for method details and validation in traditional cell culturing platforms. Methods Attempt 1 – Overstimulated cells IMR90-4 iPSC EECM-BMEC P3 frozen vial was thawed. At P4 cells were seeded and cultured 2…