human aortic vascular smcs  (Cell Applications Inc)

Journal: Bioengineering

Article Title: Mesenchymal Stem Cell-Conditioned Media-Loaded Microparticles Enhance Acute Patency in Silk-Based Vascular Grafts

doi: 10.3390/bioengineering11090947

Figure Lengend Snippet: ArtMSCs did not induce SMC toxicity. ArtMSCs and Blank-MPs were incubated with SMCs for 6, 12, and 24 h, after which the SMCs were incubated with a fluorescent LIVE/DEAD assay. Green cells indicate live cells while red cells indicate dead ones. Blank-MPs caused some cytotoxicity at 12 h and 24 h, as indicated by the red cells shown by the orange arrows. The cell death control (Dead NC using diH 2 O) and live cell control (Live PC using SBM) both indicate that the LIVE/DEAD stain was functional.

Article Snippet: Human aortic vascular smooth muscle cells (SMCs) either from ATCC (for toxicity assessment, Manassas, VA, PCS-100-012) or Cell Applications (for proliferation assays, 354-05a, San Diego, CA, USA) were cultured in supplemented basal media (SBM) (311K-500, Cell Applications Inc.).

Techniques: Incubation, Live Dead Assay, Control, Staining, Functional Assay

Journal: ACS Applied Materials & Interfaces

Article Title: Chemical Conjugation of Iron Oxide Nanoparticles for the Development of Magnetically Directable Silk Particles

doi: 10.1021/acsami.4c17536

Figure Lengend Snippet: LIVE/DEAD stain of SMCs cocultured with Silk MPs and SIMPs incubated in PBS or digested with Protease XIV showed low cytotoxicity of the particles. SMCs were also cultured with water as a negative control. Green channels (a,c,e, and g) showed confluent live cells in all MP treated groups compared to the negative control (i). Red channels (b,d,f, and h) showed minimal amounts of red (dead) cells compared to the negative control (j). The number of live (green) and dead cells (red) was counted across treatments ( n = 3), and the percentage of live and dead cells was compared to the negative control. There was a significantly higher percentage of live cells (k) and simultaneously a significantly lower percentage of dead cells (l) in the MP groups compared to the negative control. Scale bar = 100 μm ** = p < 0.01, *** = p < 0.001

Article Snippet: Human aortic vascular smooth muscle cells (SMCs) from Cell Applications (354–05a, San Diego, CA) were cultured at 37 °C and 5% CO 2 (3530, Isotemp, ThermoFisher Scientific) in supplemented basal media (311 K-500, Cell Applications Inc.) between passages 6–10.

Techniques: Staining, Incubation, Cell Culture, Negative Control

Journal: The Journal of Biological Chemistry

Article Title: Disruption of the productive encounter complex results in dysregulation of DIAPH1 activity

doi: 10.1016/j.jbc.2023.105342

Figure Lengend Snippet: Formation of a productive encounter complex increases DIAPH1-actin colocalization and regulates RAGE ligand–induced cellular migration in human vascular smooth muscle cells. A , fluorescence microscopy of HEK293T cells with GFP-tagged WT DIAPH1 ( top ), DIAPH1 ΔDAD ( middle ), and DIAPH1 E326G/E327A ( bottom ) in green , and actin labeled with Alexa Fluor 568 Phalloidin in red . Yellow indicates colocalization. B , manders coefficients, M1 and M2, quantifying colocalization between F-actin and DIAPH1 constructs and DIAPH1 constructs and F-actin, respectively. Larger values correspond to a higher degree of colocalization. Column heights reported on the graph represent mean values, error bars represent the SD. C , percent migration of human primary aortic vascular smooth muscle cells, SMCs, when stimulated with CML-HSA as compared to those treated with serum-free medium (SFM). Cells expressing YFP and DIAPH1-YFP were used as a reference and compared with those expressing DIAPH1 ΔDAD -YFP and DIAPH1 E326G/E327A -YFP. Stars indicate statistical significance: p -value <0.05 (∗), p -value <0.01 (∗∗), and p -value <0.001 (∗∗∗). D , proposed model for the equilibrium of WT DIAPH1 dimers between the active, encounter, and autoinhibited complexes. Double headed arrow indicates IH flexibility, and unstructured DAD is shown in the circle . Colored domains match that of Figure 1 A . In mouse DIAPH1, DD, CC, and FH2 domains facilitate dimerization ( , , , ). Active DIAPH1 forms a productive encounter complex via electrostatic steering, dotted line , between the RRKR motif ( blue star ) and the acidic patch of DID ( red star ). The interaction increases the probability of DID-DAD helix coupled folding and binding and formation of the autoinhibited conformation. Active DIAPH1 lacking the basic RRKR motif, such as in DIAPH-(R1213X) ( , ), cannot create a productive encounter complex resulting in an equilibrium shift toward the active complex. Note that deletion of DAD, such as in DIAPH1 ΔDAD , would preclude the autoinhibition. CC, coiled-coil; CML-HAS, carboxymethyllysine human serum albumin; DAD, diaphanous autoinhibitory domain; DIAPH1, Diaphanous 1; DID, diaphanous inhibitory domain; IH, interdomain helix; RAGE, receptor for advanced glycation end products; YFP, yellow fluorescent protein.

Article Snippet: Human primary aortic vascular SMCs (ATCC PCS-100-012) were purchased from American Type Tissue Culture Collection.

Techniques: Migration, Fluorescence, Microscopy, Labeling, Construct, Expressing, Binding Assay