Journal: Advanced healthcare materials

Article Title: Designing Inherently Photodegradable Cell-Adhesive Hydrogels for 3D Cell Culture.

doi: 10.1002/adhm.202100632

Figure Lengend Snippet: Figure 2. A) Live/dead staining of normal human dermal fibroblasts (NHDF) with calcein-AM (green, live cells) and propidium iodide (red, dead cells) embedded in GelMA-free PEGMA/PEGDMA (Gel-N0) and GelMA-containing (Gel-N10) hydrogels on day 1, 7, 14 postencapsulation, followed by imaging with confocal microscopy (Leica TCS SPE, scale bar: 200 µm). B-i) NHDF embedded in Gel-N10 hydrogels on day 1 postencapsulation. Comparison of HepG2 cells encapsulated in (ii) Gel-N10 and (iii) Gel-N0 hydrogels (scale bar: 100 µm). C) Proliferation behavior of encapsulated HepG2 cells in Gel-N10 and Gel-N0 hydrogels over a period of 7 d assessed by PrestoBlue assay (n = 3). D) Toxicity evaluation of Gel-N10 photodegradation products by MTT proliferation assay with HeLa cells after 72 h exposure (n = 3). The concentration of nondiluted degradation products was 53 mg mL−1. Data are presented as mean ± SD and statistically evaluated by (C) Student’s t-test and (D) one-way ANOVA. *, **, *** represent p < 0.05, 0.01, and 0.001, respectively.

Article Snippet: Normal human dermal fibroblasts (NHDF), human liver cancer cells (HepG2), and HeLa cells were purchased from PromoCell GmbH (Heidelberg, Germany).

Techniques: Staining, Imaging, Confocal Microscopy, Comparison, Prestoblue Assay, Proliferation Assay, Concentration Assay

Journal: BMJ Open Ophthalmology

Article Title: Protective effects of blue light-blocking shades on phototoxicity in human ocular surface cells

doi: 10.1136/bmjophth-2018-000217

Figure Lengend Snippet: Representative fluorescent microscopic images of immunostained cells with anti-pan-keratin antibody and its isotype control antibody (mouse IgG1). (A, B) Cells from human corneal surface origin (peripheral portion) and normal human dermal fibroblasts immunostained with anti-pan-keratin antibody, respectively. (C, D) Cells from human corneal surface origin (peripheral portion) and normal human dermal fibroblasts immunostained with an isotype control antibody (mouse IgG1), respectively. Scale bar=100 µm.

Article Snippet: Normal human dermal fibroblasts (NHDF, DS Pharma Biomedical, Osaka, Japan) were used as a negative control.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Hypothermically Stored Amnion Is Robust and Provides a Scaffold for Supporting Wound Healing by Retaining the Characteristics of Native Tissue

doi: 10.3390/ijms251910347

Figure Lengend Snippet: Fibroblast attachment and proliferation following seeding with human dermal fibroblasts (NHDF) and mouse fibroblasts (L929) onto hypothermically stored amniotic membranes. Cell number from AlamarBlue assessment showing metabolic activity of NHDFs ( A ) and L929s ( B ). ( C ) Representative immunofluorescence staining of seeded and non-seeded scaffolds at day 3 and 14; 40× magnification. Average ± standard deviation reported; ns denotes not significant; * denotes p < 0.05; ** denotes p < 0.01; *** denotes p < 0.001; **** denotes p < 0.0001 compared to non-seeded controls. Blue: nuclei; green: TGF-β1; red: f-actin (phalloidin); arrow: epithelial layer; S: spongy layer; Scale bar: 25 µm.

Article Snippet: Primary normal adult human dermal fibroblasts (NHDFs, lot 22TL073035 [female, age 46, Black], Lonza Biosciences, Walkersville, MD, USA) and two lots of primary mouse fibroblasts (L929, lots 14A015/70058860, ATCC, Bethesda, MD, USA) were thawed and cultured as per the manufacturer’s recommendations.

Techniques: Activity Assay, Immunofluorescence, Staining, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Hypothermically Stored Amnion Is Robust and Provides a Scaffold for Supporting Wound Healing by Retaining the Characteristics of Native Tissue

doi: 10.3390/ijms251910347

Figure Lengend Snippet: Structural characterization of fibroblasts seeded onto hypothermically stored amniotic membranes. Representative scanning electron microscopy images of non-seeded and human fibroblasts seeded ( A ) and mouse fibroblast seeded ( B ) grafts. For SEM, representative images are at 100× (scale bar: 500 µm) and 500× (scale bar: 100 µm).

Article Snippet: Primary normal adult human dermal fibroblasts (NHDFs, lot 22TL073035 [female, age 46, Black], Lonza Biosciences, Walkersville, MD, USA) and two lots of primary mouse fibroblasts (L929, lots 14A015/70058860, ATCC, Bethesda, MD, USA) were thawed and cultured as per the manufacturer’s recommendations.

Techniques: Electron Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Effect of Different Wavelengths of Laser Irradiation on the Skin Cells

doi: 10.3390/ijms22052437

Figure Lengend Snippet: Blue light (450–495 nm) effects on skin cells.

Article Snippet: Fushimi et al. [ ] , 518 nm, 0.2 J/cm 2 , human normal epidermal keratinocyte (HaCaT), primary human skin dermal fibroblasts , In fibroblasts: increased mRNA expression of HGF, KGF, leptin, IL-8 and VEGF-A increased protein levels of HGF, KGF, IL-8, VEGF-A) In keratinocytes: stimulated migration of HaCat over 24 h increased mRNA expression of HB-EGF and VEGF-A increased protein levels of HB-EGF and VEGF-A.

Techniques: Expressing, In Vivo, Isolation, Infection, Control, In Vitro, Migration, Ex Vivo, Activity Assay, DNA Synthesis, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Effect of Different Wavelengths of Laser Irradiation on the Skin Cells

doi: 10.3390/ijms22052437

Figure Lengend Snippet: Green light (495–570 nm) effect on skin cells.

Article Snippet: Fushimi et al. [ ] , 518 nm, 0.2 J/cm 2 , human normal epidermal keratinocyte (HaCaT), primary human skin dermal fibroblasts , In fibroblasts: increased mRNA expression of HGF, KGF, leptin, IL-8 and VEGF-A increased protein levels of HGF, KGF, IL-8, VEGF-A) In keratinocytes: stimulated migration of HaCat over 24 h increased mRNA expression of HB-EGF and VEGF-A increased protein levels of HB-EGF and VEGF-A.

Techniques: Expressing, Migration

Journal: International Journal of Molecular Sciences

Article Title: Effect of Different Wavelengths of Laser Irradiation on the Skin Cells

doi: 10.3390/ijms22052437

Figure Lengend Snippet: Red light (620–740 nm) effects on skin cells.

Article Snippet: Fushimi et al. [ ] , 518 nm, 0.2 J/cm 2 , human normal epidermal keratinocyte (HaCaT), primary human skin dermal fibroblasts , In fibroblasts: increased mRNA expression of HGF, KGF, leptin, IL-8 and VEGF-A increased protein levels of HGF, KGF, IL-8, VEGF-A) In keratinocytes: stimulated migration of HaCat over 24 h increased mRNA expression of HB-EGF and VEGF-A increased protein levels of HB-EGF and VEGF-A.

Techniques: Migration, Irradiation, Expressing, Membrane, Activation Assay, Co-Culture Assay, DNA Synthesis, Isolation, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Effect of Different Wavelengths of Laser Irradiation on the Skin Cells

doi: 10.3390/ijms22052437

Figure Lengend Snippet: IR light (780 nm–1mm) effects on skin cells.

Article Snippet: Fushimi et al. [ ] , 518 nm, 0.2 J/cm 2 , human normal epidermal keratinocyte (HaCaT), primary human skin dermal fibroblasts , In fibroblasts: increased mRNA expression of HGF, KGF, leptin, IL-8 and VEGF-A increased protein levels of HGF, KGF, IL-8, VEGF-A) In keratinocytes: stimulated migration of HaCat over 24 h increased mRNA expression of HB-EGF and VEGF-A increased protein levels of HB-EGF and VEGF-A.

Techniques: Isolation, Irradiation, Generated, Activity Assay, Expressing, Bacteria

Journal: Frontiers in Genetics

Article Title: Distinct Cell Stress Responses Induced by ATP Restriction in Quiescent Human Fibroblasts

doi: 10.3389/fgene.2016.00171

Figure Lengend Snippet: Effects of ATP restriction on gene regulatory networks in quiescent fibroblasts. The transcription factor (TF) network was constructed using the most enriched TFs motifs (JASPAR 2014 database). Co-regulatory connections for these motifs are provided by DNaseI footprinting (see text). Represented are the most enriched ( p < 0.005, coded in red) and avoided ( p > 0.998, coded in green) motifs, whereby p -values indicate the likelihood for a motif to occur against the background of 263 promoters in all known genes. Enriched motifs indicate a highly connected network (20 nodes, 127 edges), with many highly connected nodes including the NF-κB TF binding motif NFKB1. The network of avoided TF motifs is only sparsely connected (20 nodes, 16 links). The differentiation of the motifs by energy restriction predicts a role of Akt signaling in shaping changes in gene expression. Akt connects to TFs in the enriched group (dotted line, orange), such as EGR1, KLF4, and NFKB1. TFs known to be inhibited by Akt include FOXO TFs, HNF1A, and JUN (dotted line, neon green).

Article Snippet: The human fibroblast culture (AG10803, Coriell Institute for Medical Research, Camden, NJ) used in this study was derived from a 2 mm punch biopsy taken from the abdomen of a young donor.

Techniques: Construct, Footprinting, Binding Assay, Gene Expression

Journal: Frontiers in Genetics

Article Title: Distinct Cell Stress Responses Induced by ATP Restriction in Quiescent Human Fibroblasts

doi: 10.3389/fgene.2016.00171

Figure Lengend Snippet: NF-κBp65 binding activity levels are inversely correlated with intracellular ATP levels in quiescent fibroblasts. ATP concentrations were reduced following treatment of mitochondrial uncouplers (DNP, FCCP) and/or glycolysis inhibitor (2DG) at different concentrations as indicated. NF-κBp65 DNA binding activity was moderately increased following separate treatment with mitochondrial uncoupler (DNP/FCCP) or glycolysis inhibitor (2DG), or low concentrations of combined inhibitors. Marked increase in NF-κBp65 DNA binding activity occurred for combined treatment with mitochondrial uncouplers (DNP/FCCP) and glycolysis inhibitor (2DG) at higher concentrations. Results shown represent means and standard deviations from three independent experiments, and the best fit between ATP concentrations and NF-κBp65 DNA binding activity is an exponential function (NF-κB = 3.52 e -1.43ATP , R 2 = 0.86).

Article Snippet: The human fibroblast culture (AG10803, Coriell Institute for Medical Research, Camden, NJ) used in this study was derived from a 2 mm punch biopsy taken from the abdomen of a young donor.

Techniques: Binding Assay, Activity Assay

Journal: Cells

Article Title: Impact of Progerin Expression on Adipogenesis in Hutchinson—Gilford Progeria Skin-Derived Precursor Cells

doi: 10.3390/cells10071598

Figure Lengend Snippet: Isolation of SKPs from control and HGPS fibroblasts. ( a ) Panel showing the protocol for SKP isolation. Briefly, fibroblasts were pelleted and treated with HBSS buffer (pH 5.7) for 30 min at 37 °C. Cells were cultured in SKP media containing DMEM low glucose, EGF, FGF, and B27. The flasks were agitated daily, and the spheroids were harvested at day 4 for analysis. ( b ) SKP formation from both control (GMO1651c, GMO5565) and HGPS (HGADFN127, HGADFN003) fibroblasts with 5 and 30% senescence (SNS). ( c , d ) Quantification of the number and the diameter of the spheroids from control and HGPS fibroblast cultures with 5 and 30% SNS at day 4. Values are presented as mean ± SD ( n = 3), not significant (ns), * p < 0.05, ( c , d ) unpaired t -test. HBSS: Hank’s Balanced Salt Solution, DMEM: Dulbecco′s modified Eagle medium, EGF: epidermal growth factor, FGF: fibroblast growth factor, SKPs: skin-derived precursor cells, SNS: senescence.

Article Snippet: The human primary dermal fibroblast cell lines, GMO5565 (3-year-old male), GMO1651 (13-year-old female) and GMO5567A (12-year-old male) were all obtained from the Coriell Institute for Medical Research (Camden, NJ, USA).

Techniques: Isolation, Control, Cell Culture, Modification, Derivative Assay