Journal: STAR Protocols

Article Title: Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility

doi: 10.1016/j.xpro.2019.100013

Figure Lengend Snippet: Mechanical Rigidity of ICAM1 Nano-Topographies Controls T Cell On-Ridge Spreading and In-Groove Invasiveness Plasticity Balance (A) 3D super-resolution reconstruction of the ICAM1-functionalized PAA nano-topographic surface (G’=16 kPa). (B) Test super-resolution imaging (3D reconstruction) of human CD4+ T cells spreading and migrating along the ICAM1-coated nano-topographic surfaces on soft (16 kPa) and rigid (50 kPa) PAA surfaces. (C) Schematic of the T cell morphometric analysis and metrics: T cell height, projected area of spreading of entire cell interface ( S entire cell IF ) and projected area of in-groove invasive T cell interface ( S invasive IF ). (D and E) Mechanoregulation of T cell height, spreading area and invasiveness as indicated by T cell spreading assay on soft (G’=16 kPa) and rigid (G’=50 kPa) ICAM1 nano-textures. T cell spreading enhances on the rigid ICAM1, accompanied with T cell flattening, i.e. decrease of the T cell height. Results indicate a mechanically controlled dynamic balance between on-ridge T cell spreading and in-groove invasiveness, as shown on the schematic panel (E). I.e. T cell in-groove invasiveness structurally competes with on-ridge spreading, indicating that on-ridge spreading is mechanically enhanced and out-balances in-groove invasiveness on the rigid (G’=50 kPa) ICAM1 nano-topography. Alternatively, soft (G’=16 kPa) ICAM-1 nano-textures are unable to promote the mechanically sensitive on-ridge T cell spreading, shifting the balance towards steric in-groove T cell invasiveness. Data on the plots on (D) are as follows: boxes - means, Q1 and Q3; whiskers - max and min, X - medians; p values - one way ANOVA test. Experimental data collected in triplicates, total n>50.

Article Snippet: ICAM1, Human Protein, Recombinant, hIgG1-Fc.His Tag, Active , Sino Biological, China , Cat#10346-H03H-50.

Techniques: Imaging

Journal: STAR Protocols

Article Title: Engineering Elastic Nano- and Micro-Patterns and Textures for Directed Cell Motility

doi: 10.1016/j.xpro.2019.100013

Figure Lengend Snippet:

Article Snippet: ICAM1, Human Protein, Recombinant, hIgG1-Fc.His Tag, Active , Sino Biological, China , Cat#10346-H03H-50.

Techniques: Recombinant, Electrophoresis, Concentration Assay, Labeling, Saline, Modification, Cell Isolation, Derivative Assay, Software

Journal: International Journal of Cardiology. Heart & Vasculature

Article Title: Soluble CD54 induces human endothelial cells ex vivo expansion useful for cardiovascular regeneration and tissue engineering application

doi: 10.1016/j.ijcha.2015.01.004

Figure Lengend Snippet: Cell density of endothelial cells cultivated in the presence of soluble CD54. Endothelial cells cultivated in the presence of soluble CD54 (sCD54) were expanded for five splits. After the first (p1), second (p2) and fifth (p5) passages (as shown in the x-axis) the cellular densities were measured (y-axis) and compared between different growing conditions: in the presence of sCD54 alone, resulting in higher propagation rate, than cell culture treated with HUVEC-conditioned medium collected at p4 (CMP4) alone.

Article Snippet: Recombinant human ICAM-1/CD54 (murine myeloma cell line, NSO-derived; Accession #CAA30051), CD146, KDR and CD31 were purchased from R&D Systems, Inc.; CD45 was purchased from AbD Serotec, CD34 and CD14 were purchased from Becton Dickinson Pharmingen, and fibronectin was purchased from Southern Biotech.

Techniques: Cell Culture

Journal: International Journal of Cardiology. Heart & Vasculature

Article Title: Soluble CD54 induces human endothelial cells ex vivo expansion useful for cardiovascular regeneration and tissue engineering application

doi: 10.1016/j.ijcha.2015.01.004

Figure Lengend Snippet: Capillary density. Tube formation occurred through an ordered sequence of events and was investigated with an inverted optical light microscope; (A) cells beginning to migrate and align themselves to close polygons beginning (B) to form complete tubules (C) of endothelial cells cultivated with the addition of sCD54. The histogram shows the data mean ± SD of the quantitative analysis of tube formation area; in the x-axis was reported capillary density in the presence of HUVEC-conditioned medium collected at p4 (CMP4), sCD54 and antibody direct against CD54 at the concentration of 15 ng/ml (y-axis). Scale bars: 200 μm.

Article Snippet: Recombinant human ICAM-1/CD54 (murine myeloma cell line, NSO-derived; Accession #CAA30051), CD146, KDR and CD31 were purchased from R&D Systems, Inc.; CD45 was purchased from AbD Serotec, CD34 and CD14 were purchased from Becton Dickinson Pharmingen, and fibronectin was purchased from Southern Biotech.

Techniques: Sequencing, Light Microscopy, Concentration Assay

Journal: Frontiers in Immunology

Article Title: A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

doi: 10.3389/fimmu.2023.1276151

Figure Lengend Snippet: Upon exposure to inflammatory stimuli such as LPS, or sensitizing chemicals such as 1 chloro-2,4-dinitrobenzene (DNCB) or nickel sulfate (NiSO 4 ), keratinocytes start to secrete inflammatory cytokines such as IL-1, TNF-α and IL-18. Subsequently cutaneous dendritic cells such as Langerhans cells and dermal dendritic cells become activated and start to phagocytose haptens or exogenous particles, which is accompanied by cell maturation and the upregulation of CD54 and CD86. Finally, DCs migrate to draining lymph nodes to present the processed antigen in order to activate CD4 + T cells. Created with BioRender.com .

Article Snippet: For staining the cells were incubated in Automacs Running Buffer with the following antibodies (1:50): REA Control (S)-VioGreen (Miltenyi, #130-113-444), REA Control (S)-PE (Miltenyi, #130-113-438), REA Control (S)-APC (Miltenyi, #130-113-434); REA Control (S)-PE-Vio770, (Miltenyi, #130-113-440); CD54-APC (Miltenyi, #130-121-342); CD86-APC (Miltenyi, #130-116-161), CD14-VioGreen (Miltenyi, #130-110-525), CD11c-APC (Miltenyi, #130-113-584) for 10 minutes in the dark.

Techniques:

Journal: Frontiers in Immunology

Article Title: A human 3D immune competent full-thickness skin model mimicking dermal dendritic cell activation

doi: 10.3389/fimmu.2023.1276151

Figure Lengend Snippet: Surface marker expression of CD54 and CD86 (depicted as fold of induction of the percentage of all positive cells) after (A) pre-treatment of THP-1-derived iDCs and (B) topical treatment of the immune competent skin model with dexamethasone for 1 h, followed by NiSO 4 treatment for 23 h. Results were depicted as fold of induction compared to the solvent control [0.3% DMSO]. (C–E) Cytokine secretion of iDCs after 1 h dexamethasone pre-treatment, followed by 23 h of NiSO 4 exposure. Error bars indicate the standard errors of the mean (n = 3 independent experiments for (A, C) and n=4 independent experiments for (B) with *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, and ****p ≤ 0.0001).

Article Snippet: For staining the cells were incubated in Automacs Running Buffer with the following antibodies (1:50): REA Control (S)-VioGreen (Miltenyi, #130-113-444), REA Control (S)-PE (Miltenyi, #130-113-438), REA Control (S)-APC (Miltenyi, #130-113-434); REA Control (S)-PE-Vio770, (Miltenyi, #130-113-440); CD54-APC (Miltenyi, #130-121-342); CD86-APC (Miltenyi, #130-116-161), CD14-VioGreen (Miltenyi, #130-110-525), CD11c-APC (Miltenyi, #130-113-584) for 10 minutes in the dark.

Techniques: Marker, Expressing, Derivative Assay, Solvent, Control