Journal: Nutrients

Article Title: Essential Oils, Pituranthos chloranthus and Teucrium ramosissimum , Chemosensitize Resistant Human Uterine Sarcoma MES-SA/Dx5 Cells to Doxorubicin by Inducing Apoptosis and Targeting P-Glycoprotein

doi: 10.3390/nu13051719

Figure Lengend Snippet: Effects of Pituranthos chloranthus (PC) and Teucrium ramosissimum Desf. (TR) extracts on normal primary human uterine fibroblast cells (HUF) and primary murine Bone Marrow-Derived Macrophages (BMDM) viability. After treatment of primary HUF and murine BMDM with increasing concentrations (0–100 µg/mL) of PC and TR for 72 h, the percentage of viable cells was assessed using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ( A ) Dose–response curves of PC-treated HUF (left panel) and TR-treated HUF (right panel). ( B ) Dose–response curves of PC-treated BMDM (left panel) and TR-treated BMDM (right panel). Data are expressed as a mean percentage of control growth ± Standard Deviation (SD) of two representative experiments ( n = 6 replicates per concentration).

Article Snippet: Primary human uterine fibroblast normal cells (HUF) were obtained from the ATCC.

Techniques: Derivative Assay, MTT Assay, Control, Standard Deviation, Concentration Assay

Journal: Allergy, asthma & immunology research

Article Title: Serum Amyloid A1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients.

doi: 10.4168/aair.2022.14.1.40

Figure Lengend Snippet: Fig. 1. The production of SAA1 from AECs and the effects of SAA1 on proinflammatory cytokine production. SAA1 proteins were produced from AECs (SAECs and A549, respectively) in response to various stimuli (A, B), and in the coculture with PBNs, PBEs (C, D). SAA1-induced release of IL-6, IL-8 and S100A9 from AECs (SAEC and A549, respectively) (E-K). Data are presented as means ± standard error of mean (n = 6). P values were calculated by using one-way analysis of variance with Bonferroni's post hoc test. SAA1 enhanced the expression of p-ERK and p-p38 MAPK (L) in AECs (SAECs and A549, respectively). IL, interleukin; PBEs, peripheral blood eosinophils; PBNs, peripheral blood neutrophils; dex, dexamethasone; S100A9, S100 calcium-binding protein A9; p-ERK, phospho-extracellular signal-regulated kinase; ERK, extracellular signal-regulated kinase; p-p38 MAPK, phospho-p38 mitogen-activated protein kinase; MAPK, mitogen-activated protein kinase; AECs, airway epithelial cells; SAEC, small airway epithelial cells. *P < 0.050, †P < 0.010, ‡P < 0.001, ns = not significant.

Article Snippet: Epithelial cell culture and stimulation Two kinds of AECs were used in this study, primary small airway epithelial cells (SAECs) and human AEC line (A549), both of which were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Produced, Expressing, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: High-Content Imaging and Machine Learning Classify Phenotypical Change in Coronary Artery Endothelial Cells Caused by BPS

doi: 10.3390/ijms27073259

Figure Lengend Snippet: Representative high-content microscopy images of human coronary artery endothelial cells (HCAEC) exposed to vehicle control (CTRL) or 0.1 µM Bisphenol S (BPS) for 96 h and stained using the PhenoVue Cell Painting assay. For each condition, a representative field acquired at 40× magnification and a higher-magnification inset are shown. Rows correspond to the individual fluorescence channels: Hoechst 33342 (nuclei), PhenoVue Fluor 488 Concanavalin A (endoplasmic reticulum and intracellular membranes), PhenoVue 512 nucleic acid stain (RNA/nucleoli), PhenoVue Fluor 555 wheat germ agglutinin (plasma membrane), PhenoVue 641 mitochondrial stain (mitochondria), and the merged image. White boxes represent the part of the image used for the related inset. Scale bar: 50 µm, 40× objective.

Article Snippet: Primary human coronary artery endothelial cells (HCAEC; ATCC ® PCS-100-020TM, Innovation, VA, USA) were cultured according to the supplier’s recommendations.

Techniques: Microscopy, Control, Staining, Fluorescence, Clinical Proteomics, Membrane

Journal: Cancers

Article Title: Preclinical Evaluation of the Novel Small-Molecule MSI-N1014 for Treating Drug-Resistant Colon Cancer via the LGR5/β-catenin/miR-142-3p Network and Reducing Cancer-Associated Fibroblast Transformation

doi: 10.3390/cancers12061590

Figure Lengend Snippet: Cancer-associated fibroblasts (CAFs) increased the oncogenic properties of colon cancer cells. ( A ) Insert illustrates the co-culture system of DLD1 and HCT116 colorectal cancer (CRC) cells with Normal fibroblasts (NF) and CAFs. Cell viability assay showed increased 5-fluorouracil (5-FU) resistance in CAF-educated DLD1 and HCT116 cells, compared to their naïve counterparts. Flow cytometric analysis of DLD1 and HCT116 cells co-cultured with CAFs. Increased CD44 + cell population ( B ) and side population ( C ) in both DLD1 and HCT116 cells, 48 h post CAF culture. Enhanced colony-forming ( D ), migratory ( E ), and tumor sphere-generating ( F ) abilities in both CRC cell lines post CAF co-culture. ** p < 0.01, *** p < 0.001.

Article Snippet: The DLD1 and HCT116 human colon cancer cell lines and normal fibroblasts (NFs, ATCC® PCS-201-018) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured according to the vendor’s recommended conditions.

Techniques: Co-Culture Assay, Viability Assay, Cell Culture

Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 1 Harnessing fibroblast contractility to engineer dense carcinoma constructs. (a) Schematic illustration of fibroblast‐driven contraction of collagen fibers around cancer spheroids. Blue shapes represent fibroblasts, black lines represent collagen fibers, and green clusters represent spheroids. (b) Side by side comparison of H&E‐stained sections of dense breast carcinoma constructs (left) and biopsied mammary ductal carcinoma tissue (right). Scale bars = 80 μm. (c) Time‐course imaging of gross contraction and compaction of dense carcinoma constructs. Scale bars = 200 μm. (d) Box plot of the relative area change over time for gels seeded at 2.5 × 105 HLF/mL (n = 6) and 5 × 105/mL (n = 7). **p < 0.01. ***p < 0.001. See Section 2 for details of statistical analyses. (e) Box plot of the corresponding increase in estimated collagen density over time for gels seeded for acellular gels and 5 × 105 HLF/mL (n = 5 per group).

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Construct, Comparison, Staining, Imaging

Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 2 Effect of fibroblast cell type and seeding density on tissue contraction. (a–c) Time‐course imaging of gross contraction and compaction of dense carcinoma constructs formed using several types of fibroblasts. Constructs were seeded at 5 × 105/mL (n = 3 per group). Scale bars = 3 mm. Similar kinetics and magnitude of contraction using human dermal fibroblasts (HDF, a), human lung cancer associated fibroblasts (CAF, b), and human lung fibroblasts (HLF, c). (d and e) Time‐course imaging of gross contraction and compaction for up to 3 weeks in culture. Scale bars = 3 mm. Constructs were seeded at 0.5 × 105 HLF/mL (n = 6, panel d) and 5 × 105/mL (n = 7, e). (f) Relative area change over time as a measure of contraction for each type of fibroblast during 7 days of culture as shown in (a–c). No significant differences were measured. HDF constructs were slightly less contracted at 7 days, matching the qualitatively larger size in time course images. (g) Relative area change over time as a measure of contraction. Similar final volumes were reached with each seeding density. Contraction was more gradual with a seeding density of 0.5 × 105 HLF/mL.

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Imaging, Construct

Journal: Biotechnology and bioengineering

Article Title: Engineering dense tumor constructs via cellular contraction of extracellular matrix hydrogels.

doi: 10.1002/bit.28561

Figure Lengend Snippet: FIGURE 5 Local anisotropy is similar in constructs formed with various types of contractile cells. (a) H&E‐staining of sectioned dense carcinoma constructs formed with cancer associated fibroblasts (CAF), human dermal fibroblasts (HDF), and human lung fibroblasts (HLF). Seven days, scale bars = 30 μm. (b) Collagen type I staining of sectioned dense carcinoma constructs formed with CAF, HDF, and HLF. Seven days, scale bars = 20 μm. (c) Box plot of the relative fluorescent intensity in binned regions of interest at the specified distances from spheroid for dense carcinoma constructs formed with CAF (white), HDF (light gray), and HLF (dark gray) (n = 9 per group). *p < 0.05, **p < 0.01. One way ANOVA. (d) Box plot of fiber orientation relative to spheroid tangent line and distance from spheroid for dense carcinoma constructs. Same shading as (d) to indicate fibroblast type; CAF (white), HDF (light gray), and HLF (dark gray) (n = 9 per group). One way ANOVA. ns, not significant.

Article Snippet: Normal human lung fibroblasts (HLF) (American Type Culture Collection [ATCC] and Lonza) were cultured in fibroblast basal medium with low serum kit (ATCC) and 2% fetal bovine serum (FBS, ATCC).

Techniques: Construct, Staining

Journal: Scientific Reports

Article Title: A tool kit for rapid cloning and expression of recombinant antibodies

doi: 10.1038/srep05885

Figure Lengend Snippet: (A–B) Flow cytometric histograms showing fluorescent intensity of cells incubated with purified IgG/IgE antibodies plus secondary FITC-labelled goat anti-human IgG/IgE (open) and cells with secondary antibody only (filled) against the number of cells. (A) Flow cytometric assessment of anti-CSPG4 IgG 1 /IgE antibodies shows specific binding to native CSPG4 antigen present on the cell surface of A375 melanoma cells and no binding above background to primary melanocytes. (B) Fc regions of anti-CSPG4 IgG 1 and 102.1F10 IgG 4 isotypes demonstrate effector-binding to U937 monocytic cell line, expressing human Fcγ receptors. The IgE antibody isotypes also bind similarly to RBL SX38 mast cells, expressing human FcεR1 receptor. (C) Immunofluorescence staining of A375 cells confirms specific binding of anti-CSPG4 IgG 1 /IgE and no background binding with isotype control hapten specific NIP-IgG 1 /IgE detected by goat anti-human IgG/IgE-FITC. (D) Grass pollen allergen specificity is confirmed by anti-human sandwich ELISA, showing specific binding of recombinant 102.1F10 IgG 4 /IgE and original patient serum to the ELISA plate-bound Phl p 7 allergen and no binding above background with unspecific human myeloma IgG 4 and MOv18 IgE isotype controls.

Article Snippet: Primary human epidermal melanocytes (PCS-200-012, ATCC) were grownin Dermal Cell Basal Medium (PCS-200-030, ATCC) and supplemented with the Melanocyte Growth Kit (PCS-200-041, ATCC).

Techniques: Incubation, Purification, Binding Assay, Expressing, Immunofluorescence, Staining, Control, Sandwich ELISA, Recombinant, Enzyme-linked Immunosorbent Assay