Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Detection of the OSMR on lung fibroblasts. Cells were stained with no antibody or with anti-OSMR followed by FITC-labelled goat anti-rabbit IgG. Immunostaining was then analysed by flow cytometry.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Staining, Immunostaining, Flow Cytometry

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Proliferation of human lung fibroblasts in response to OSM. Cells were exposed to OSM at concentrations ranging from 0.5 – 100 ng ml−1 for 24, 48 or 72 h. Proliferative effect was examined using an MTS assay (A) and confirmed by [3H]-TdR uptake (B). The proliferative effect of OSM was completely abrogated by incubation of cells with neutralizing antibodies against OSMR and gp 130. Results are means of four separate experiments performed in triplicate. *P<0.05 compared to cells in serum free conditions.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: MTS Assay, Incubation

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Cell counts of human lung fibroblasts after incubation with OSM

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Incubation

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Detection of cyclin D1 expression in lung fibroblasts. Immunohistochemical staining for cyclin D1 was performed on unstimulated cells (D) or cells exposed to OSM (2 ng ml−1) for 24 h (E). The specificity of the cyclin D1 antibody was confirmed by substituting the primary antibody with non-immune mouse IgG1 (F). Cells were also stained with DAPI in order to enumerate cell nuclei (A – C). Immunofluorescent images from 1 μm sections were obtained by scanning laser confocal microscopy at 40×magnification.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Expressing, Immunohistochemical staining, Staining, Confocal Microscopy

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Effects of the tyrosine kinase inhibitor (genestein) and p42/44 MAPK inhibitor (PD98059) on OSM-induced proliferation. Fibroblasts were seeded in 24 well plates and incubated with genestein (10 μM) or PD98059 (50 μM) in the presence or in the absence of 2 ng ml−1 OSM. At 24 and 48 h cell numbers were assessed using an MTS assay. Results are means of four separate experiments performed in triplicate. *P<0.05 versus cells exposed to OSM 2 ng ml−1. #P<0.05 compared to 24 h time point.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Incubation, MTS Assay

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Effects of the non-selective COX inhibitor indomethacin and the selective COX-2 inhibitor nimesulide on OSM-induced proliferation, COX expression and PGE2 release. (a) Cells were treated with indomethacin (5 μM) or the selective COX-2 inhibitor nimesulide (5 μM) prior to incubation with OSM (2 ng ml−1). Proliferative responses to OSM were not influenced by either drug at 24 or 48 h. Detection of COX-1 and COX-2 protein. Fibroblasts were incubated for either 6 or 24 h with OSM (2 ng ml−1) or IL-1β (2 ng ml−1) and expression of both COX-1 and COX-2 was determined by Western analysis (b). PGE2 release in response to OSM. PGE2 release was measured by EIA in supernatants taken from fibroblasts treated with OSM (2 ng ml−1) for 24 and 48 h (c). Results are shown as means of four separate experiments. *P<0.05 compared to cells in serum free medium.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Expressing, Incubation, Western Blot

Journal:

Article Title: Oncostatin M stimulates proliferation, induces collagen production and inhibits apoptosis of human lung fibroblasts

doi: 10.1038/sj.bjp.0704769

Figure Lengend Snippet: Effect of OSM on human lung fibroblast pro-collagen production. Fibroblasts were grown to confluence and incubated with 10 or 100 ng ml−1 OSM for 48 h and the production of hyp measured by HPLC. Data are means of six independent experiments. *P<0.05 compared to cells in serum free medium.

Article Snippet: The normal diploid human foetal lung fibroblast cell line was obtained from American Type Culture Collection (ATCC, Rockville, MD, U.S.A) and was used between passages 11 and 24.

Techniques: Incubation

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: Genes to Cells

Article Title: Derivation of human differential photoreceptor cells from adult human dermal fibroblasts by defined combinations of CRX, RAX , OTX2 and NEUROD

doi: 10.1111/gtc.12127

Figure Lengend Snippet: Induction of retina-specific genes in human dermal fibroblasts by the retroviral infection of genes for defined transcription factors. (A) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (NHDF) obtained from Lonza after gene transfer of several kinds of transcription factors. Recoverin, blue opsin and PDE6c genes were up-regulated by CRN transduction. ‘Negative control’: amplified water as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘w/o’: cultured fibroblasts without gene transfer as the other negative control. ‘SPPO’: SOX2, POU1F1, PAX6 and OTX2 . ‘SPO’: SOX2, POU1F1 and OTX2 . ‘CRN’: CRX, RAX and NEUROD . ‘Human retina’: human retinal tissue as a positive control. The amount of cDNA as a template was a half in the positive control. (B) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (NHDF) obtained from Promo Cell after gene transfer of several transcription factors. Recoverin, blue opsin and PDE6c genes were up-regulated by CRN or CRNO transduction. ‘w/o’: cultured fibroblasts without gene transfer as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘CRNO’: CRX, RAX, NEUROD and OTX2 . (C) RT-PCR analysis for photoreceptor-specific genes in cultured human dermal fibroblasts (HDF-a) obtained from ScienCell after gene transfer of several transcription factors. Recoverin, blue opsin and PDE6C genes were up-regulated by CRN or CRNO transduction. Expression levels of blue opsin were increased by additional OTX2 gene transduction. ‘w/o’: cultured fibroblasts without gene transfer as a negative control. ‘GFP’: cultured fibroblasts after retroviral gene transfer of the GFP gene as another negative control. ‘CRNO’: CRX , RAX , NEUROD and OTX2 . ‘1’, ‘2’ and ‘3’ mean independently cultured, transfected and harvested cells by the same combination of CRN genes. (D) Immunocytochemistry using antibodies to rhodopsin and blue opsin (green). Nuclei were stained with DAPI (blue). Experiments were carried out at 2 weeks after infection. The cells in the left panel and the right panel are CRN-infected Fib#2 and Fib#1, respectively. Scale bars represent 10 μm.

Article Snippet: Three strains of cultured human dermal fibroblasts were used: one was obtained from Lonza (NHDF), another was from Promo Cell (NHDF) and the other was from ScienCell (HDF-a).

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Transduction, Negative Control, Amplification, Positive Control, Expressing, Transfection, Immunocytochemistry, Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) Pretreatment with low dose ActD decreases the proportion of 8N cells caused by VX-680 in NHDF cells. ActD was added 24 hours post-seeding. VX-680 was added 48 hours post-seeding. Cells were harvested 96 hours after seeding. (B) NHDFs were treated with 4 nM ActD for 72 hours, fixed and stained with Giemsa (top panel). In the bottom panel, ActD was removed from the medium and cells were allowed to proliferate for 6 days before Giemsa staining.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A and B) NHDF cells were treated for 48 hours with VX-680 alone (labelled as no preinc.), or treated for 48 hours with VX-680 after preincubating with the indicated amounts of ActD for 24 hours. Thereafter cells were fixed, stained Giemsa (A) and counted (B). Arrows in (A) indicate cells with abnormal nuclei. (C and D) NHDF cells treated as in A and B were allowed to recover for 6 days in drug-free medium. The growth of the cultures in drug-free medium is quantified in panel (C) by counting the total number of cells per field before and after the recovery period. The average number of cells per field with aberrant nuclei before and after the recovery period is shown in panel (D). Error bars correspond to standard deviations.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) NHDF cells were treated with the indicated amounts of ActD for 4 or 12 hours. p53 and p21 were analysed by Western blotting. α-tubulin was detected as a loading control. (B) Example of nuclear abnormalities in NHDF cells expressing a dominant negative form of p53. The percentage of cells with this sort of aberrant nuclei was 2.5 fold higher than in the corresponding cells with intact p53. Severely damaged cells, such as the one in the center of this picture, only appeared in the fibroblasts with dominant negative p53.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Western Blot, Expressing, Dominant Negative Mutation

Journal:

Article Title: ?-Melanocyte-Stimulating Hormone Counteracts the Suppressive Effect of UVB on Nrf2 and Nrf-Dependent Gene Expression in Human Skin

doi: 10.1210/en.2008-1315

Figure Lengend Snippet: Expression analysis of Nrf1-3 in human skin cells in vitro and in situ. A, Detection of Nrf1-3 in various human skin cell types in vitro as shown by conventional RT-PCR analysis. NC, Negative control, H2O as template; 1, NHK; 2, HaCaT; 3, NHM; 4, HDF; 5, HMEC-1. B, Protein expression of Nrf2 in human skin cell types. Total cell lysates (50 μg/lane) were subjected to Western immunoblot analysis using an anti-Nrf2 antibody. The same blot was reprobed with an anti-α-tubulin antibody. C, Expression of Nrf2 in healthy human skin. Nrf2 was detected by immunohistochemistry using the immunoperoxidase technique (a). Note specific cytoplasmic Nrf2 immunoreactivity within the basal layer of epidermal keratinocytes (red cells, arrows) but not in the IgG control (c). Double immunostaining of Nrf2 and melanocytes by the immunoperoxidase (Nrf2) and immunogold-silver enhancement technique (melanocytes) using a pan-Mel antibody (b). Note gray-blackish precipitate within a melanocyte but lack of Nrf2 staining within (arrow).

Article Snippet: Cell culture Normal human keratinocytes (NHK) and human dermal fibroblasts (HDF) were all derived from newborn foreskin and purchased from PromoCell (Heidelberg, Germany).

Techniques: Expressing, In Vitro, In Situ, Reverse Transcription Polymerase Chain Reaction, Negative Control, Western Blot, Immunohistochemistry, Double Immunostaining, Staining