Last meeting, we discussed the possibility of the storage gel-boxes outgassing a residue onto the porous SiO2 membranes, as evidenced by cells not adhering to the membranes and difficulty in acquiring SEM images. Possible Gel-Box Outgassing on Porous SiO2 Membranes Several cleaning methods were utilized in attempt to clean the membranes, such as O2 plasma,…
In previous experiments attempting to culture ADSC on non-porous and porous (0.5 µm, 3.0 µm) SiO2 membranes, we were having difficulty getting the ADSC to attach to the membranes. A majority of the ADSC did not appear well-spread. While in culture or staining, ADSC would detach from the membrane with little to no agitation. Changes…
YAP/TAZ YAP/TAZ intracellular localization, and subsequent transcriptional activity, has indicated its role in pathways of mechanotransduction. Investigation of the effects of substrate stiffness on MSC growth and differentiation point to YAP/TAZ as an indicator of how cells are perceiving their mechanical microenvironment and a regulator of the cells’ response to this environment. MSCs cultured on stiff…
Objective Develop an easily mainpulated reproducible vasculogenesis assay using hydrogels laden with endothelial cells (‘bio-ink’) patterned using the technique of extrusion bioprinting Bio-inks Develop a bio-ink with the following characteristics Cell viable: bio-ink must sustain cell life, allowing cells to manipulate and move through the hydrogel Vasculogenic: bio-ink must encourage (or at least not negatively affect) tubule formation…
Background: The purpose of these experiments was to investigate the formation of gap junctions between co-cultured HUVEC and ADSC across 0.5µm HP SiO2 and 0.4 µm HP Track-Etched membranes. Gap junctions are channels that allow small molecules to pass directly from cell to cell. Electron micrographs show that the membranes of two adjacent cells are…
To supplement our upcoming manuscript, we decided that we needed to provide a visual of how thin our ultrathin SiO2 membranes actually are. To demonstrate the thinness of the SiO2 membranes, in comparison to the commercially available Track-Etched hanging membrane inserts, we performed a co-culture using 0.5µm high porosity SiO2 membranes (thickness ~0.1-0.3µm) and 0.5µm…
Introduction Permeable membranes provide a physiologically relevant cell culture substrate for in vitro study of barrier cell types, such as endothelial cells, which assume a polarized morphology with distinct apical and basolateral surfaces. Cell morphology and subsequent monolayer formation and barrier function are largely dependent on initial interactions with the cell culture substrate via focal adhesions….
The following post is the abstract that I recently submitted for the BMES Annual Conference. Introduction: Permeable membranes provide a physiologically relevant cell culture substrate for in vitro study of barrier cell types, such as endothelial cells, which assume a polarized morphology with distinct apical and basolateral surfaces. Cell morphology and subsequent monolayer formation and…
Purpose: To investigate the formation of focal adhesions (FAs) on various cell culture substrates, including TCP, SiO2 membranes, and PDMS membranes. Background: FAs are large macromolecular assemblies through which mechanical force and regulatory signals are transmitted between the extracellular matrix (ECM) and an interacting cell. FAs consist of a complex network of ECM, membranes, and cytoplasmic components…
Background: Similar to the Gaborski lab, the authors of the publication “Microfabrication of high-resolution porous membranes for cell culture” fabricated and characterized ultra-thin polymeric membranes for use as cell culture substrates. Upon reviewing their publication, Tom and I discussed some of the experiments that we’d like to replicate to further characterize the SiO2 membrane for our…