For everyone conducting biological studies, this paper highlights how to think about sample size and replicates. It suggests using “SuperPlots”, which color codes data points from different experiments to better visualize the type of variability seen (i.e. random variability, experimental…
Author: Molly McCloskey
Localizing Basement Membrane Production in Coculture BBB Model
Introduction The basement membrane (BM) is a critical component of a healthy blood-brain barrier (BBB) model. Further, the BM breaks down in several diseases. Both pericytes and endothelial cells can contribute to basement membrane production. This post will explore a…
Optimizing IMR90-4 Coculture BBB Model
Introduction While we have optimized coculture models for commercial brain endothelial cells and pericytes, we needed to do the same for iPSC-derived cocultures. In this post, I show the optimization process using IMR90-4-derived EECM-BMEC-like cells [1] and brain pericyte-like cells (BPLC)…
Posted in NRG
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…
Developing a Distributable µSiM ‘Transwell’-like Platform…Publication Figure Drafts
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…
IMR90-4 EECM-BMEC-like Cell Analysis in µSiMs
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…
Posted in memBrains
in situ Permeability Analytical Solution Summary – Unhindered vs Hindered D
Introduction Pages regarding our in situ permeability assay development can be found in these links: Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 1: Approach) Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 2: Experimental Validation)…
Posted in memBrains
Sampling Method Permeability Assay Optimization
Motivation We have discussed the optimization of our in situ permeability assay at length. This assay relies on confocal microscopy, which may not be accessible to all users of the µSiM. Therefore, we developed an alternative permeability assay in the…
hiPSC Differentiation to Endothelial Cell Culture Method, Brain Microvascular Endothelial Cells (EECM-BMEC-like cells), Round 2
Introduction This post highlights results from my second attempt to differentiate hiPSCs into endothelial progenitor cells EECM-BMEC-like cells in our lab. For details on methodology, see Nishihara et al., FASEB, 2020 and my previous post. Methods Cells were seeded at 70,000 and 100,000…
Posted in NRG
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…
