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: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) A schematic outlining the strategy for SILAC-based identification of SASP proteins. See for time lines of how cells were made quiescent or senescent and how quiescent and senescent conditioned media (CM) were produced. (B) Reactome pathway analysis of proteins secreted at >2-fold levels by senescent compared with quiescent HCA2 cells. Pathways related to hemostasis are identified by boxes. (C) Heatmap showing individual levels of proteins, identified by SILACs, that participate in hemostasis and are secreted by quiescent (QUI) and senescent (SEN [IR]) HCA2 normal human fibroblasts. (D) CM were collected from quiescent (QUI) and senescent (SEN [IR]) cells, concentrated, and analyzed using western blotting for the indicated SASP factors. Cells remaining on the plate at the time of CM collection were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. Fold changes are mean ± SEM. *p < 0.05 and **p < 0.01. See also and .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Multiplex sample analysis, Produced, Western Blot, Staining

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet: (A) mRNA was extracted from HCA2 cells made quiescent (QUI) or induced to senesce by doxorubicin (SEN [DOXO]), and analyzed using qPCR for the indicated genes, using actin mRNA for normalization. The data are shown in a.u., and the bars show mean ± SEM. *p < 0.05 and **p < 0.01, two-tailed, unpaired t test. (B) CM were collected from cells treated as in (A), concentrated and analyzed using western blotting for the indicated SASP factors. Cells were counted for normalization. Mindin protein levels and Ponceau S staining were used as loading controls. (C) IMR-90 fibroblasts were depleted of mtDNA by culture in ethidium bromide until senescent (21 days) (labeled as MiDAS). RNA was extracted and analyzed using qPCR for the indicated factors. Heatmap represents individual values for three experiments. *p < 0.05, one-way ANOVA. See also .

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Two Tailed Test, Western Blot, Staining, Labeling

Journal: Cell reports

Article Title: SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells

doi: 10.1016/j.celrep.2019.08.049

Figure Lengend Snippet:

Article Snippet: WI-38 and IMR-90 are normal primary human embryonic lung fibroblasts (obtained from the American Type Culture Collection).

Techniques: Recombinant, Modification, Sequencing, Multiplex sample analysis, Labeling, Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Reverse Transcription, Software

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: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 1. Effects of specific wavelengths on the ability of IL-1α to stimulate matrix-related mRNA in fibroblasts. Human cultured fibroblasts were irradiated with 30 mJ of UVB, 5 J of UVA, 5 J of UVA1, or sham irradiated, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, followed by 24 h of incubation at 37°C. Displayed are normalized levels of tropoelastin (A) and collagenase (B) mRNA.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Cell Culture, Irradiation, Incubation

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 2. Effects of specific wavelengths on cellular secretion of TNF-α. (a) Fibroblasts were irradiated with 30 mJ of UVB, 5 J of UVA, 5 J of UVA1, or sham irradiated, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, followed by 24 h of incubation at 37°C. Displayed are TNF-α concentrations in the conditioned media (n 5 2). (b) Keratinocytes were similarly irradiated, incubated, and media assayed for TNF-α. Displayed are TNF-α concentrations in the conditioned media when no exogenous IL-1α had been added (n 5 2). Separate experiments demonstrated that exogenous IL-1α did not affect secretion of TNF-α by keratinocytes exposed to any of these wavelengths. (c) Neonatal and adult keratinocytes were irradiated with 10 or 30 mJ per cm2 of UVB or 5 or 20 J per cm2 of UVA. Displayed are TNF-α concentrations in the conditioned media from irradiated neonatal (n 5 2) and adult (n 5 3) keratinocytes, p , 0.01 for adult 30 mJ per cm2 UVB relative to sham by analysis of variance, then Dunnett q9 statistic.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Irradiation, Incubation

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 3. Effects of exogenous TNF-α on the ability of IL-1α to stimulate tropoelastin mRNA in fibroblasts. Human cultured fibroblasts were incubated with or without IL-1α and TNF-α, as indicated. Displayed are normalized tropoelastin mRNA levels after 24 h of incubation at 37°C. IL-1α produced a statistically significant increase in tropoelastin mRNA only in the cells that received no TNF-α (p , 0.01 by t test).

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Cell Culture, Incubation, Produced

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 4. Effect of anti-TNF-α antibodies on the ability of UVB to block the effect of IL-1α on matrix-related messages. Human cultured fibroblasts were irradiated with 30 mJ of UVB or sham irradiation, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, plus the indicated concentrations of anti-TNF-α inhibitory antibodies. Displayed are normalized tropoelastin (a) and collagen (b) mRNA levels after 24 h of incubation at 37°C.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Blocking Assay, Cell Culture, Irradiation, Incubation