Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: SM induces human dendritic cell maturation which is further enhanced by cancer cell necroptosis (A) Schematic representation of the in vitro co-culture assay. moDCs alone (B) or in co-culture with spheroids (tumor cells and hd-fibroblasts) (D and E) were treated with the compound conditions depicted. 72 h post-treatment, cells were harvested and analyzed via flow cytometry. (B) Relative proportion of activated vs non-activated moDC treated alone. (C) TSNE plot depicting islands of phenotypically similar cells based on CD45, CD11c, HLA-DR, CD86, CD83, and PDL1 as markers. Included are the moDCs treated in monoculture, as well as the different co-cultures. Analyzed cells separate into three subpopulations: activated moDC – blue, non-activated moDC - light gray, tumor cells - black. (D) Relative proportion of tumor and moDCs in the different co-cultures. (E) Relative proportion of activated vs non-activated moDCs in co-culture. Error bars, mean ± SD of triplicate independent wells for each condition of at least 2 independent experiments using different donors. ∗∗p value <0.01, by two-tailed t-test. TNFα: 0.1 ng/mL, SM: 1 μM, zVad: 5 μM. (F) Heatmap depicting cytokines measured in the supernatant of spheroids composed of tumor cells with or without CAFs, 72 h post-treatment. Data are averages from 3 independent wells and black squares indicate levels measured below the detection limit. SM: 0.25-1 μM; TNFα: 0.1 ng/mL, zVad: 20 μM.

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: In Vitro, Co-culture Assay, Co-Culture Assay, Flow Cytometry, Two Tailed Test

Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: SM modulates fibroblasts by downregulating myofibroblast-like CAF markers and upregulating pro-inflammatory soluble mediators CAF 2D monoculture was treated with increasing concentrations of SM or TGFβ. (A) Fold change in FAP and αSMA expression, analyzed via flow cytometry, 72 h post-treatment. (B) CAF confluence monitored overtime using IncuCyte. Error bars, mean ± SD of 3 independent experiments. (C) Heatmap depicting fold change in cytokine and chemokine concentration in the supernatant of treated CAFs (normalized to control). Results from 2 independent experiments. Black squares indicate levels measured below the detection limit.

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: Expressing, Flow Cytometry, Concentration Assay, Control

Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: Key resources table

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software, Simple Western, High Content Screening

Journal: Acta Dermato-Venereologica

Article Title: Novel Detection and Clinical Utility of Serum-Derived Extracellular Vesicle in Angiosarcoma

doi: 10.2340/actadv.v105.40902

Figure Lengend Snippet: miR-3926, miR-5703, miR-3925-5p, and miR-184 were upregulated in angiosarcoma (AS) cell lines. Expression of microRNAs (miRNAs) in an AS cell line (ISO-HAS B) and benign human endothelial cell lines (HUVEC and HDMVECn.) were compared by quantitative reverse transcription PCR. ISO-HAS B showed significant upregulation of miR-5703 ( P adj = 0.0078 and P adj = 0.0261, respectively) and miR-184 ( P adj = 0.0214 and P adj = 0.0053, respectively) in comparison with HUVEC and HDMVECn (F value for each one-way ANOVA analysis was 12.14 and 14.06, respectively). ISO-HAS B also showed a tendency to upregulations in miR-3926 and miR-3925-5p, while the differences were not significant (F value for each one-way ANOVA analysis was 3.154 and 3.237, respectively). The experiment was performed 3 times with triplicate samples for each cell line. HUVEC: human umbilical vein endothelial cell; HDMVECn: human dermal microvascular endothelial cells neonatal. * p < 0.05, ** p < 0.01.

Article Snippet: Commercial human umbilical vein endothelial cells (HUVEC) (ATCC ® , Manassas, VA, USA, PCS-100-010TM) and human dermal microvascular endothelial cells neonatal (HDMVECn) (ATCC ® , PCS-110-010TM) were cultured in vascular cell basal medium (ATCC ® , PCS-110-030TM) supplemented with an Endothelial Cell Growth Kit-VEGF (ATCC ® , PCS-110-041TM).

Techniques: Expressing, Reverse Transcription, Comparison

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Schematic depicting the protocol for hCECs, hCSSCs, and hiNSCs seeding onto silk scaffolds.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques:

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Evaluating the effects of the microenvironment (elevated glucose or dynamic perfusion) and different chemical stimulants (capsaicin and sodium lauryl sulfate) on cell viability and gene and protein expression using the corneal tissue model. ( Abbreviations : Interleukin-1 beta (IL-1β), Substance P (SP), Calcitonin gene-related peptide (CGRP); Gene names : CRCP -accessory protein for CGRP, SCN- sodium channel, KERA - keratocan (cornea-specific proteoglycan), LUM - lumican (proteoglycan abundant in the stroma)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Expressing, Concentration Assay, Permeability

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Common secreted proteins related to nerve activation or tissue inflammation present in conditioned media isolated from corneal constructs. The relative concentration ranges of each protein detected in conditioned media in response to various chemical stimuli have been previously reported by our group ( Deardorff et al., 2018 ; Siran et al., 2018 ). ( Abbreviations : Substance P (SP), Calcitonin gene-related peptide (CGRP), Interleukin-1 beta (IL-1β), Tumor necrosis factor-alpha (TNF-α), Matrix metalloproteinase-9 (MMP-9)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Activation Assay, Isolation, Construct, Concentration Assay

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Phenotypic markers of each cell type present in the corneal construct. ( Abbreviations : Transient receptor potential cation channel subfamily V member 1 (TRPV1), Transient receptor potential cation channel subfamily A member 1 (TRPA1), Transient receptor potential cation channel subfamily M member 8 (TRPM8)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Marker

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Troubleshooting guide for construct assembly and characterization.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Cell Culture, Microscopy, Incubation, Sterility, Isolation, Lysis, Homogenization, Binding Assay

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Relative timeframe for completion of each step in the assembly, stimulation, and characterization of the corneal tissue model. The amount of time may vary significantly depending on the user and the total number of constructs generated. These timeframes are based on 12 constructs per experiment.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Generated, Incubation, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay, Imaging, Lysis, Protein Concentration, BIA-KA, Western Blot

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Structure and in vitro activities of BKI-1748 against N. caninum (Nc-β-gal) and T. gondii (Tg-β-gal). (A) molecular structure of BKI-1748, MW = molecular weight; EC 50 = concentration of half maximal proliferation inhibition; *toxicity levels against human foreskin fibroblasts (HFF) and zebrafish embryos were determined previously [22]. (B,C) Dose-response curves for Neospora and Toxoplasma tachyzoites grown in HFF.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: In Vitro, Molecular Weight, Concentration Assay, Inhibition

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Litter size, parasite burden, neonatal and postnatal mortality rates of N. caninum infected mice treated with BKI-1748.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: Infection