Nanomembrane Research Group

Protocols (Public)

Immunofluorescence staining of junctional proteins and adhesion molecules of hiPSC-derived EECM-BMEC-like cells on nanomembranes
ByPelin Kasap February 23, 2021June 7, 2023

I would like to share my successful immunofluorescence stainings of adhesion molecules and junctional proteins of hiPSC-derived brain microvascular endothelial cell (BMEC)-like cells on nanomembranes. The cells were differentiated by the extended endothelial cell culture method (EECM). You can find detailed information about EECM-BMEC-like cells in the previous post submitted by Hideaki Nishihara (24.10.2020). Method…

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hiPSC Differentiation into Brain Pericyte-like Cells, Round 2
ByMolly McCloskey February 22, 2021February 25, 2021

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…

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HBVP (pericyte) and hCMEC/D3 (endothelial cell) µSiM Coculture Optimization
ByMolly McCloskey February 20, 2021April 28, 2021

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…

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iPSC IMR90-4 ApoE Genotyping
ByMolly McCloskey February 20, 2021February 20, 2021

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…

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AFM studies on HUVECs in Aline Devices
ByWilliam Houlihan January 27, 2021

Introduction Previous work was done on determining physical properties of the glycocalyx of endothelial cells, this was performed on glass inserts using the Asylum AFM. We wanted to continue this testing using the Aline uSim devices in order to determine any significant differences between our devices and the glass bottom flow system developed in…

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Theoretical Underpinnings of Small Molecule Permeability Measurements in the µSiM (Part 1: Approach)
ByJim January 27, 2021February 23, 2021

NOTE: Some of the ideas in this post are borrowed and rephrased from prior posts by Molly and Alec and our paper Khire et al. 2020. The purpose of this thread is to explain things anew with an eye on application in the R61 project. Small molecule diffusion from the well to the channel side…

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hToC Modular Approach
ByRaquel Ajalik January 26, 2021

Background: The human Tendon-on-Chip (hToC) spans collaboration between the Center for Musculoskeletal Research (CMSR) at the UR and the McGrath lab to develop an in vitro platform to study inflammation, fibrosis, and cell-cycle regulation in fibrovascular tendon healing. Interaction between the blood-tendon-barrier (BTB) is fundamental to the investigation of this pathology as part of the…

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Multiplexed digital assay: mouse CCL2(MCP1) CXCL1(KC) panel development
ByHoward Su January 26, 2021January 26, 2021

Mouse CCL2/MCP1 and CXCL1/KC pre-equilibrium digital ELISA has been developed with the current dynamic range set to be 4-500pg/ml for CCL2/MCP1 and 8-1000pg/ml for CXCL1/KC. This initial dynamic range was selected by inferring from the ELISA kit reagent working concentration given by the vendor R&D systems (1.9-250pg/ml for CCL2/MCP1 & 8-1000pg/ml for CXCL1/KC) and can…

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ALine mod-µSiM: Version2
ByMolly McCloskey January 24, 2021January 25, 2021

Modular µSiM Version 2 (V2) Holds ~30 µl of media in the bottom channel. Designed for cocultures, ELISAs, and other assays that are easier with greater channel media volumes. For more surface area and volume information on mod-µSiM V2, see ALine Modular Device Geometry. Passive Pumping Another feature of V2 devices is their ability to…

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Flow demo (T cell interaction with human in vitro BBB model under flow)
ByPelin Kasap January 19, 2021

20210122.flow-movie-demo I attached a link to a short presentation regarding the flow movie setups. The figures and videos will be explained in detail during the meeting on Friday.

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