Author: Barrett Nehilla

Images of 1st BBB co-culture (+ 2 extras)

This post presents cell images from the first successful co-culture experiment.  I seeded P8 bEnd3 cells alone, P13 NG108-15 cells alone and both cells on PET and SC500 pnc-Si transwells.  For this experiment, I seeded 50,000 bEnd3/cm2 on the apical

Posted in NRG

Investigating vacuoles on transwells

One of the themes in my recent posts has been the presence of ‘vacuoles’ over free-standing pnc-Si and PET membranes. We’ve assumed these are vacuoles because endothelial cells form vacuoles, they don’t stain with live cell stains (and so have

Posted in NRG

Endothelial actin networks on pnc-Si transwells

I’ve been wondering if there were differences in actin/cytoskeletal structure of cells on supported vs. free-standing pnc-Si.  A couple weeks ago, I had a couple of extra pnc-Si transwells so I stained for actin. I followed the actin staining (rhodamine-phalloidin)

Posted in NRG

H2O2 crosses "nonporous" membranes

A couple weeks ago, I showed some preliminary data on H2O2 detection via the Amplex Red Assay, which is a project for one of our high school students.  This generated some discussion at the last NRG meeting about diffusion through

Posted in NRG

(Very) small molecule diffusion through pnc-Si transwells

We have a high school student, Nikita, (from my Summer Nanotech class) who is volunteering in the lab.  Jim and I have assigned him the project of exploring diffusion of very small molecules through porous and putatively nonporous membranes.  Currently,

Posted in NRG

Endothelial cells form vacuoles within 1 day on pnc-Si

In this experiment, I wanted to test how quickly vacuoles form in endothelial cells when cultured on pnc-Si transwells.  Here I used 30nm SC502 RTP’ed samples with the pnc-Si side down.  I seeded P9 bEnd3 cells at 50000 cells/cm2 on

Posted in NRG

Porosity vs. Cell Morphology

As I was preparing for the BMES conference, I took a few pictures of endothelial cells on pnc-Si transwells that showed extensive vacuole/endosome expression (here).  The most dramatic example occurred on a 50nm-thick membrane that was barely porous.  So I

Posted in NRG

Glial cells can grow as multilayers upside-down

In my 1st post detailing glial cells on pnc-Si transwells, I saw lots of cell clumps above the membrane window (see here).  Since these cells were grown in the wells, I thought they could be piling on top of each

Posted in NRG

Side-dependent barrier function

I’ve suspected that the barrier function of endothelial cells might be surface-dependent (that is, cells grown on the pnc-Si side vs. cells grown on the well side).  I hypothesized that the interface between membrane and well walls would alter the

Posted in NRG

Monolayers, not clumps on pnc-Si membrane

In my last couple of posts with respect to endothelial cell culture on pnc-Si transwells, I presented images of cell clumps over the active pnc-Si membrane area of transwells (here and here).  There were dead cells associated with these clumps

Posted in NRG
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