In my last post, I presented the results of my BBB TEER experiments. For the BBB co-culture, there was a spike in TEER. I hypothesized that this was caused by media acidification by rapidly proliferating glial cells, which in turn negatively affected barrier function of endothelial cells. When I presented this data to Angela Glading,…
This post presents the data from all of my TEER experiments of BBB co-cultures, with bEnd3 endothelial cells on the bottom and NG101815 glia on the top of PET and pnc-Si transwells. For all experiments, I used 50000 cells/cm2 as the seeding density. I always had 3 samples – bEnd3 alone, NG10815 alone and the…
A few weeks ago, I posted some high resolution (100X) images of BBB co-cultures stained with 3 different colors. I also included the z-height of the different focal planes of cells on either side of PET transwells. In this post, I present a repeat of this experiment, along with the same analysis performed with cells…
To follow up my last post on air-blood barrier TEER results, I wanted to show some Live/Dead images of these samples. These are stained after the 14 day experiment. In the following image, you can see normal bEnd morphology on PET and pnc-Si. Only the cells over the pnc-Si slit are visible because these cells…
I finished 3 independent experiments of air-blood barrier co-cultures – these consist of bEnd3 endothelial cells on 1 side and RLE-6TN lung epithelial cells on the opposite side of the membranes. We didn’t expect these cell types to potentiate TEER in anyway (like we expected glial cells to tighten endothelial barrier function). However, our collaborators…
Hi everyone! I’m sifting through the mass of data I collected during my last month and am compiling posts to update everyone. This post shows the latest with the microfluidic chamber that Henry and I designed. In my first post regarding this device, I showed microparticle and blood flow over cultured endothelial cells (bEnd3). In…
Over the last couple of weeks, I’ve been doing confocal fluorescence microscopy over at the Med Ctr. I was hoping this tool would clean up some of the background fluorescence in my immunofluorescent images. More importantly, the confocal can take ‘slices’ of images in the z-plane and stack them on top of each other to…
As a proof-of-principle, Henry and I have been working on a microfluidic device for flowing blood cells/particles across cultured cells on pnc-Si. More on why this is important later. We’ve come up with a prototype and I’ve been testing it out. I will include schematics later but here are the first couple of movies I’ve…
One of the points I’d like to discuss in my co-culture paper is the advantageous imaging qualities of pnc-Si compared to ‘translucent’ commercial transwell membranes. This has been evident in many of my posts of Live/Dead stained cells, even at 20X. I thought a higher resolution (100X) image would emphasize this point even more since…
I’ve been trying to get multi-color images of endothelial cells and glial cells on transwell membranes in order to compare image quality on glass, pnc-Si and polymer membranes. Specifically, I’ve tried staining endothelial cells green with VEGF mAbs and glial cells red with neurofulament H (NFH) mAbs but VEGF expression in bEnd3 cells is extremely…