I am trying to grow HUVECs under flow in my device to establish a tight and confluent monolayer for TEER analysis. My device schematic looks like this and the whole setup is as below I tried to use the NPN chip in sepcon format so that I don’t have to be restricted by number of…
In conducting experiments to address reviewer comments, we again created endothelial-like cells from adipose derived stem cells (ADSCs). As in the past, the vast majority of ADSCs express some CD31 (PECAM-1) after culture in endothelial media (M200) supplemented with 50ng/mL VEGF (on 0.5LP, 3.0LP and TCP). The expression of this CD31 is relatively low and…
This is part of our work studying how cells adhere, spread and proliferate on varying pore size/porosity SiO2 membranes. This post will be a qualitative look at how HUVEC spread over membranes by staining for actin filaments. In a future post we will quantify the degree of spreading at 1hr, 4hrs and 24 hrs. Calculation…
Barrett had discovered that growing endothelial cells on pnc-si or tranwells causes vacuoles to form. The vacuoles on pnc-si formed only on the free standing areas of the membrane where there are open pores. The background on pnc-si chips showed negligible vacuole formation. The density of vacuoles on pnc-si was almost two fold higher than…
This experiment was carried out to determine the maximum life of pnc-Si in cell culture conditions based on discoloration. All the discoloration slowing factors were put together to see how long would the membranes last for. These include: samples in 3D format (Sepcon) with ‘membrane side’ down (with no intact membranes), all in incubator, DMEM…
After four days of this ongoing HUVEC culture in pncSi transwell, HUVECs look much better than before. Here I have freshly stained them with Calcein AM and we can see how bright it is in comparison to CMFDA stained cells (or may be there’s some flaw in my CMFDA staining procedure). Till now its been…
On Day 2 of cell growth from trial 3 of adhesion study, cells looked much more spread out but less in density. Most of the cells were observed to have concentrated near the edge of the pnc-Si chip near the Sepcon as seen in the figure below (first image in the bottom row). This could…
Samples (no RTP) from W673 and W684 were taken (four from each wafer; with atleast one blank chip) in Sepcons and HUVECs were plated @ ~19,000 cells/sq. cm. density. Cells were allowed to adhere for over 4 hours and this time stained with CMFDA (instead of Calcein). Cell types loaded with the CellTracker probes are…
According to recent cell adhesion studies on aminosilanized pncSi, no significant difference between adherence to the silanized and non-silanized surfaces was found. This could have been due to the adsorption of protein from the cell growth media onto the surface nullifying the effect due to silanization which is supposed to make it hydrophobic. ‘If silane…
I repeated the previous experiment, this time plating slightly higher density of cells: ~15100 cells/sq.cm. Samples were from the same W673, no RTP) After counting the cells stained using Calcein AM, % Adhesion w.r.t the cells plated were found to be: non-silanized samples = 55.87%; silanized samples = 41.33% In above figures, not many cells…