First off, this is the protocol for the staining and analyzing the data is shown below and was used for all of the YAP experiments: For the substrates, we wanted to use 0.5μm HP, 0.5μm LP, 3μm HP, TCP, and nonporous SiO2. Cyto-Vu devices were made using the aforementioned substrates and 300 ADSCs were seeded per membrane. The…
To follow up on my recent posts about HUVEC YAP localization after 24 and 72 hours (https://trace-bmps.org/blog/data/2016/10/19/yaptaz-passage-one-staining and https://trace-bmps.org/blog/data/2016/10/11/yaptaz-staining-on-membranes-and-nonporous-sio2) , we conducted the same study using Adipose Derived Stem Cells. Due to the fact that these are mesenchymal stem cells, we thought that their expression of YAP might be a bit different due to the…
After reading and discussing a paper in journal club that talked about cells having a “mechanical memory” (found here “Mechanical memory and dosing influence stem cell fate“), we decided to replicate our previous YAP/TAZ experiments with passage one HUVECs direct from Life Technologies as to avoid making our cells “remember” that they had ever been plated…
As we’re uncovering more bits and pieces of data, we kept being lead to the idea that our membranes may be made out of SiO2 and have a modulus in the gigapascal range, but cells seem to perceive them as soft substrates. After looking at focal adhesions, fibronectin deposition, and cell/cell junction formation, we’re still sticking…
To continue our spreading and proliferation experiments on various topographical features and stiffness, I repeated the HUVEC experiments I had done before due to a possible artificial bias that we think is a result of high seeding. To enusre accuracy, I repeated the experiment and added another plate to test the HUVECs on substrates of…
To analyze the dimpled Sylgard substrates that we’ve been making, we’ve been conducting experiments to investigate how topographical differences in the surface of a substrate impacts HUVECs and ADSCs, in this case the dimples. We’ve already shown that cells will grow on these membranes in our “Dimpled Sylgard Membrane Device” post and that our substrates are wetting in our “Degassing Experiments”…
As we were conducting our experiments on dimpled Sylgard substrates we knew that another micro-topography that we wanted to investigate was micro-pillars. In like manner to the dimples, a post microtopography should provide some sort of surface for the cells to “grab onto” and spread as they would on any ECM in vivo. These…