Cruciferous vegetable-derived indole-3-carbinol prevents coronavirus cell egression mechanisms in tracheal and intestinal 3D in vitro models.

Generation of Porcine and Rainbow Trout 3D Intestinal Models and Their Use to Investigate Astaxanthin Effects In Vitro

“…This is confirmed by the results in Figures 1C and 2C, which show that the formation of the stromal and the epithelial compartments in both the porcine and rainbow trout species can be appreciated. The results are also consistent with previous observations that describe an Alvetex ® thickness of 200 µm [42] as an ideal feature to replicate in vitro the distance of cells from blood capillaries, thus replicating the behavior of nutrients in vivo [41], and is in line with the choice of several authors that previously adopted this approach to generate body barriers in vitro, such as the intestinal mucosa [43,45], the dermis [41,42,46], and the endometrium [47,48]. Notably, all of these studies demonstrated the development of the two main barrier compartments, namely the stroma and the epithelium, similar to what is described in the present manuscript and suggested the ability of Alvetex ® scaffold to encourage epithelial-ECM interactions, which typically occur in vivo [49,50].…”

“…We then take advantage of the newly created functional 3D models to investigate whether AST may interact with the molecular mechanisms possibly involved in inflammation and oxidative stress. In particular, in the present experiments, 3D cell aggregation is achieved with the use of a highly porous polystyrene scaffold, commercially available and registered under the name Alvetex ® , which is able to maintain the cells’ original shape and encourage cell interactions [ 41 , 42 , 43 , 44 , 45 ]. This is confirmed by the results in Figure 1 C and Figure 2 C, which show that the formation of the stromal and the epithelial compartments in both the porcine and rainbow trout species can be appreciated.…”

“…Based on all these observations, in the present work, we used primary cells, freshly isolated from intestinal biopsies to maintain the cell’s original phenotype and, more in general, to preserve the anatomical and physiological features distinctive of the intestinal epithelial cells in vivo. As described above, in the 3D models created, the presence of the epithelial compartment is demonstrated by the active transcription of the typical tight junction markers, as well as the expression of adherens junction-related genes, suggesting the activation of a molecular expression pattern that supports barrier integrity [ 45 ]. This is in agreement with previous studies that used this strategy and showed that the formation of intercellular junctions contributed to the mechanical properties of individual epithelial cells, cell adhesion, and intercellular communication [ 56 ].…”

Single Nucleus RNA Sequencing of Remnant Kidney Biopsies and Urine Cell RNA Sequencing Reveal Cell Specific Markers of Covid-19 Acute Kidney Injury

“…Recent studies show that the HECT-E3 ligase family including genes such as NEDD4 and WWP1 interact and ubiquitylate the SARS-CoV-2 spike protein. Furthermore, the HECT family members are overexpressed in COVID-19 infected patients, where rare germline activating variants in NEDD4 and WWP1 are associated with severe COVID-19 cases [37][38][39] . Thus, the repertoire of COVID-19 AKI genes we observe represent markers that can exacerbate COVID-19 and others that are associated with countering its adverse effects.…”