primary human newborn foreskin fibroblasts nuff-1 cells  (GlobalStem)

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of human BJ fibroblasts stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining before and after 5.0 ng/mL transforming growth factor (TGF)β1 treatment for 6 days. N = 3 biological replicates. b Representative flow cytometry plot of BJ fibroblasts before and after 5.0 ng/mL TGFβ1 treatment for 6 days, with mouse anti-α-SMA antibody [4A4] (GTX60466, GeneTex; 1: 250 dilution) and goat anti-mouse IgG(H + L) highly cross-adsorbed secondary antibody, Alexa Fluor TM 405 (A-31553, Thermo Fisher Scientific; 2 μg/mL). c Quantification of α-SMA + myofibroblast percentages in BJ fibroblasts from ( b ). N = 3 biological replicates. d Expression of ACTA2 (the gene encoding α-SMA) in BJ fibroblasts before and after fibroblast-myofibroblast conversion. Data were normalized to the ACTA2 level before TGFβ1-induced fibroblast-myofibroblast conversion. N = 3 biological replicates. e Representative images of BJ fibroblast-converted myofibroblasts (BJ-myofibroblasts) with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. N = 3 (for α-SMA and TUNEL staining) or 4 (for flow cytometry) biological replicates, respectively. f Quantification of apoptotic BJ-myofibroblasts by flow cytometry from ( e ). g Quantification of pyroptotic BJ-myofibroblasts by flow cytometry from ( e ). N = 4 biological replicates ( f , g ). Scale bars, 25 μm. Data presented as mean ± s.d. overlaying all the data points. P- values were determined by two-tailed unpaired t -tests ( c – g ). N.S ., not significant, P > 0.05; * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: Human newborn foreskin BJ fibroblasts (CRL-2522 TM ; ATCC) at passage 27 were used for genomic editing and RNAi, as well as fibroblast-to-myofibroblast conversion and apoptosis analysis.

Techniques: Staining, Immunofluorescence, Flow Cytometry, Expressing, TUNEL Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of the primary NHDF FB-AA36 (derived from a 36-year-old female African–American donor; Supplementary Table ) stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining before and after transforming growth factor (TGF)β1-induced fibroblast-myofibroblast conversion. b Expression of ACTA2 in FB-AA36 fibroblasts before and after fibroblast-myofibroblast conversion. Data were normalized to the ACTA2 level before TGFβ1-induced fibroblast-myofibroblast conversion. c Representative images of NHDF FB-AA36 with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. d Quantification of NHDF FB-AA36 apoptosis from flow cytometry in ( c ). e Representative images of FB-AA36 fibroblast-converted myofibroblasts with α-SMA and TUNEL staining, accompanied by the respective flow cytometry plots stained with Annexin V-FITC and PI staining. f Quantification of FB-AA36 fibroblast-converted myofibroblast apoptosis from flow cytometry in ( e ). Scale bars, 25 μm. Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P- values were determined by two-tailed unpaired t -tests ( b – f ). N.S ., not significant, P > 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: Human newborn foreskin BJ fibroblasts (CRL-2522 TM ; ATCC) at passage 27 were used for genomic editing and RNAi, as well as fibroblast-to-myofibroblast conversion and apoptosis analysis.

Techniques: Derivative Assay, Staining, Immunofluorescence, Expressing, TUNEL Assay, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of the human keloid-derived fibroblast KB-AA35 (derived from a 35-year-old female African–American donor; Supplementary Table ) stained with α-smooth muscle actin (α-SMA) by immunofluorescence staining. Scale bar, 25 μm. b Expression of ACTA2 in FB-AA36 fibroblasts and KB-AA35 fibroblasts. Data were normalized to the ACTA2 level of FB-AA36 fibroblasts. c Quantification of KB-AA35 fibroblast apoptosis by flow cytometry with Annexin V-FITC and PI staining. d Expression of IL1B in KB-AA35 fibroblasts with or without FMOD treatment. Data were normalized to the IL1B level without FMOD treatment. e Active IL1β production in KB-AA35 fibroblasts with or without FMOD treatment. Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P- values were determined by two-tailed unpaired t -tests ( b – e ). * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: Human newborn foreskin BJ fibroblasts (CRL-2522 TM ; ATCC) at passage 27 were used for genomic editing and RNAi, as well as fibroblast-to-myofibroblast conversion and apoptosis analysis.

Techniques: Derivative Assay, Staining, Immunofluorescence, Expressing, Flow Cytometry, Two Tailed Test

Journal: Nature Communications

Article Title: Fibromodulin selectively accelerates myofibroblast apoptosis in cutaneous wounds by enhancing interleukin 1β signaling

doi: 10.1038/s41467-025-58906-z

Figure Lengend Snippet: All cells were subjected to serum starvation prior to treatment. a Representative images of normal skin (from abdomen), keloid (from abdomen), and hypertrophic scar (from left shoulder) tissues collected from the donor with the National Disease Research Interchange (NDRI) reference ID 1914138 with α-smooth muscle actin (α-SMA) staining by immunohistochemistry (IHC; upper), and the representative images of dermal fibroblasts isolated from these tissues with α-SMA staining by immunofluorescence staining (lower). b Expression of ACTA2 in dermal fibroblasts derived from tissues described from ( a ). Data were normalized to the ACTA2 level of fibroblasts isolated from the normal skin. c Representative flow cytometry plots of fibroblasts derived from tissues described in ( a ) with Annexin V-FITC and PI staining. d Quantification of apoptotic dermal fibroblasts by flow cytometry from ( c ). e The influence of FMOD on cell apoptosis with and without IL1β was calculated from ( d ). Scale bars, 50 μm (black) or 25 μm (white). Data presented as mean ± s.d. overlaying all the data points. N = 3 biological replicates; P -values were determined by two-tailed unpaired t -tests ( b – e ). N.S ., not significant, P > 0.05; * P < 0.05; ** P < 0.005. Source data are provided as a Source Data file.

Article Snippet: Human newborn foreskin BJ fibroblasts (CRL-2522 TM ; ATCC) at passage 27 were used for genomic editing and RNAi, as well as fibroblast-to-myofibroblast conversion and apoptosis analysis.

Techniques: Staining, Immunohistochemistry, Isolation, Immunofluorescence, Expressing, Derivative Assay, Flow Cytometry, Two Tailed Test

Journal: European Heart Journal Open

Article Title: Modulation of cell fate by shock wave therapy in ischaemic heart disease

doi: 10.1093/ehjopen/oeaf011

Figure Lengend Snippet: Shockwave therapy induces transdifferentiation via Toll-like receptor 3. ( A ) Expression of the innate immune receptor Toll-like receptor 3 on human cardiac fibroblasts. green = Toll-like receptor 3; blue = DAPI. Scale bar = 20 µm. ( B ) Toll-like receptor 3 activation of Toll-like receptor 3 reporter cells upon shockwave therapy or Poly(I:C) treatment (30 ng/mL). Data are means ± SEM. ** P < 0.01. ( C ) The expression of the downstream adapter TIR-domain-containing adapter-inducing interferon-β after shockwave therapy or Poly(I:C) treatment (30 ng/mL). ( D ) mRNA levels of interleukin-6 after shockwave therapy or Poly(I:C) treatment (30 ng/mL). Data are means ± SEM. * P < 0.01. ( E ) Human fibroblasts were treated with shockwave therapy in the presence of endothelial induction medium containing bone morphogenetic protein 4, vascular endothelial growth factor, and fibroblast growth factor and subjected to reverse transcription polymerase chain reaction (RT-PCR) or fluorescence-activated cell sorting. ( F ) mRNA expression of CD31 after treatment with control medium, endothelial induction medium, shockwave therapy, or a specific dsRNA/Toll-like receptor 3 inhibitor. Data are means ± SEM. ** P < 0.01. ( G ) mRNA expression of VEGF receptor 2 after treatment with control medium, endothelial induction medium, shockwave therapy, or a specific dsRNA/Toll-like receptor 3 inhibitor. Data are means ± SEM. * P < 0.05. ( H ) mRNA expression of vascular endothelial cadherin after treatment with control medium, endothelial induction medium, shockwave therapy, or a specific dsRNA/Toll-like receptor 3 inhibitor. Data are means ± SEM. ** P < 0.01. ( I ) Fluorescence-activated cell sorting for CD31 after treatment with shockwave therapy or a combined treatment with a specific dsRNA/Toll-like receptor 3 inhibitor and shockwave therapy. Statistical comparisons between multiple groups: one-way analysis of variance with Tukey post hoc analysis. ALP, alkaline phosphatase; BMP4, bone morphogenetic protein 4; CTRL, control; EIM, endothelial induction medium; FACS, fluorescence-activated cell sorting; FGF, fibroblast growth factor; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IL-6, interleukin-6; SWT, shockwave therapy; TLR3, Toll-like receptor 3; TLR3inh, TLR3 inhibitor; TRIF, TIR-domain-containing adapter-inducing interferon-β; VE-cadherin, vascular endothelial cadherin; VEGF, vascular endothelial growth factor; VEGFR2, VEGF receptor 2.

Article Snippet: BJ human newborn foreskin fibroblasts (ATCC, Washington, DC, USA) were cultured and maintained in DMEM with 10% FCS and 1% penicillin/streptomycin/glutamine at 37°C and 5% CO 2 .

Techniques: Expressing, Activation Assay, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Fluorescence, FACS, Control

Journal: European Heart Journal Open

Article Title: Modulation of cell fate by shock wave therapy in ischaemic heart disease

doi: 10.1093/ehjopen/oeaf011

Figure Lengend Snippet: Induced endothelial cells are functional in vitro and in vivo . ( A ) mRNA expression of CD31 after fluorescence-activated cell sorting of induced endothelial cells or fibroblasts. Data are means ± SEM. ** P < 0.01. ( B ) mRNA expression of vascular endothelial growth factor receptor 2 after fluorescence-activated cell sorting of induced endothelial cells or fibroblasts. Data are means ± SEM. **** P < 0.0001. ( C ) CD31 expression of induced endothelial cells or fibroblasts after fluorescence-activated cell sorting. red = CD31; blue = DAPI. Scale bar = 20 µm. ( D ) Tube formation assay with induced endothelial cells or fibroblasts after fluorescence-activated cell sorting. Scale bar = 50 µm. ( E ) Number of nodes per high power field analysed in a tube formation assay in induced endothelial cells or fibroblasts. Data are means ± SEM. **** P < 0.0001. ( F ) Number of junctions per high power field analysed in a tube formation assay in induced endothelial cells or fibroblasts. Data are means ± SEM. **** P < 0.0001. ( G ) Nitric oxide concentration produced by induced endothelial cells or fibroblasts. Data are means ± SEM. **** P < 0.0001. ( H ) Induced endothelial cells, fibroblasts or human umbilical vein endothelial cells were suspended in Matrigel and injected subcutaneously into NSG mice. Blood perfusion of the plug was measured via laser Doppler perfusion imaging (left image). The right image shows the injection sides of the plugs. ( I ) Blood perfusion of the subcutaneously injected Matrigel plugs containing fibroblasts, induced endothelial cells, or human umbilical vein endothelial cells. Data are means ± SEM. **** P < 0.0001. ( J ) Immunofluorescence staining of an induced endothelial cell containing Matrigel plug for CD31. red = CD31; blue = DAPI. Scale bar = 50 µm. ( K ) Hematoxylin and eosin staining (H.E.) sections of explanted Matrigel plugs containing fibroblasts, induced endothelial cells, or human umbilical vein endothelial cells analysed for vessels (arrowheads). scale bar = 30 µm. ( L ) Number of vessels per high power field in HE stained sections containing fibroblasts, induced endothelial cells, or human umbilical vein endothelial cells. Data are means ± SEM. * P < 0.05. Statistical comparisons between two groups: Student’s t -test, multiple groups: one-way analysis of variance with Tukey post hoc analysis. HPF, high power field; HUVEC, human umbilical vein endothelial cell; iEC, induced endothelial cell; NO, nitric oxide; VEGF, vascular endothelial growth factor; VEGFR2, VEGF receptor 2.

Article Snippet: BJ human newborn foreskin fibroblasts (ATCC, Washington, DC, USA) were cultured and maintained in DMEM with 10% FCS and 1% penicillin/streptomycin/glutamine at 37°C and 5% CO 2 .

Techniques: Functional Assay, In Vitro, In Vivo, Expressing, Fluorescence, FACS, Tube Formation Assay, Concentration Assay, Produced, Injection, Imaging, Immunofluorescence, Staining

Journal: European Heart Journal Open

Article Title: Modulation of cell fate by shock wave therapy in ischaemic heart disease

doi: 10.1093/ehjopen/oeaf011

Figure Lengend Snippet: Shockwave therapy induces transdifferentiation in ischaemic myocardium and improves cardiac function. ( A ) To test whether we could induce transdifferentiation of cardiac fibroblasts within a myocardial scar towards endothelial cells, we created a transgenic mouse model of Fsp1-Cre/LacZ mice and performed a lineage tracing experiment after ligation of the left anterior descending and shockwave therapy. ( B ) Left ventricular fibrosis assessed via Masson trichrome staining after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control). Scale bar = 1 mm. ( C ) Quantification of left ventricular fibrosis after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control). Data are means ± SEM. * P < 0.05. ( D ) M-mode images after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control). ( E ) Left ventricular ejection fraction after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control) (%). Data are means ± SEM. ** P < 0.01. ( F ) Left ventricular end-diastolic diameter after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control) (mm). Data are means ± SEM. ** P < 0.01 ( G ) Cardiac output after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control) (mL/min). Data are means ± SEM. ( H ) Immunofluorescence staining of Fsp1-Cre/LacZ mice after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control). red = CD31; green = LacZ. blue = DAPI. ( I ) Number of LacZ/CD31-positive cells after left anterior descending ligation with (shockwave therapy) or without shockwave therapy (control). Data are means ± SEM. **** P < 0.0001. Statistical comparisons between two groups: Student’s t -test, multiple groups: one-way analysis of variance with Tukey post hoc analysis. CTRL, control; HPF, high power field; LV, left ventricular; LVEDD, left ventricular end-diastolic diameter; SWT, shockwave therapy.

Article Snippet: BJ human newborn foreskin fibroblasts (ATCC, Washington, DC, USA) were cultured and maintained in DMEM with 10% FCS and 1% penicillin/streptomycin/glutamine at 37°C and 5% CO 2 .

Techniques: Transgenic Assay, Ligation, Staining, Control, Immunofluorescence

Journal: European Heart Journal Open

Article Title: Modulation of cell fate by shock wave therapy in ischaemic heart disease

doi: 10.1093/ehjopen/oeaf011

Figure Lengend Snippet: Shockwave-treated hearts show larger endothelial-like fibroblast subcluster after myocardial infarction. ( A ) Uniform manifold approximation and projection plot showing all sequenced nuclei from all samples from different time points (control Day 1, control Day 14, shockwave (SWT) therapy Day 1, and shockwave therapy Day 14). ( B ) Uniform manifold approximation and projection plot showing fibroblast subclustering, revealing 10 fibroblast subclusters (Leiden clustering, resolution 0.5). ( C ) Gene expression of endothelial and fibroblast marker genes in the different fibroblast clusters shown in B . ( D ) Bar plots showing the relative fibroblast cell fraction of fibroblast Subcluster 8 at different time points. ( E ) Gene ontology analysis of differential expressed genes in fibroblast Cluster 8 at 14 days after shockwave therapy vs. control. ( F ) Partition-based graph abstraction showing cluster dynamics among fibroblast subclusters. Edges represent cluster connectivities; threshold: 0.1). CTRL, control; SWT, shockwave therapy; TLR, Toll-like receptor; UMAP, uniform manifold approximation and projection; VEGF, vascular endothelial growth factor.

Article Snippet: BJ human newborn foreskin fibroblasts (ATCC, Washington, DC, USA) were cultured and maintained in DMEM with 10% FCS and 1% penicillin/streptomycin/glutamine at 37°C and 5% CO 2 .

Techniques: Control, Gene Expression, Marker

Journal: bioRxiv

Article Title: The Human Cytomegalovirus vGPCR UL33 is Essential for Efficient Lytic Replication in Epithelial Cells

doi: 10.1101/2024.09.18.609710

Figure Lengend Snippet: Fibroblasts (NuFF-1) were infected (MOI= 0.5 TCID50/cell) with WT, UL33-3xF, UL33Δ1, or UL33Δ2 and analyzed at 96 hpi. (A) Whole cell lysates were probed by immunoblot with the indicated antibodies. IE1, marker of lytic infection; actin, loading control. N = 3, representative blots shown. (B) Cells were fixed, permeabilized, and UL33 localization was assessed using an antibody against the FLAG epitopes (green). mCherry (red) is shown as a marker of infection, and DAPI (blue) was used to visualize nuclei. Images were acquired at 63X magnification. N= 3, representative images shown.

Article Snippet: Primary newborn human foreskin fibroblasts (NuFF-1) were obtained from GlobalStem.

Techniques: Infection, Western Blot, Marker, Control

Journal: bioRxiv

Article Title: The Human Cytomegalovirus vGPCR UL33 is Essential for Efficient Lytic Replication in Epithelial Cells

doi: 10.1101/2024.09.18.609710

Figure Lengend Snippet: (A) Fibroblasts (NuFF-1) were infected (MOI= 0.5 TCID50/cell) with TB40/E- mCherry -UL33-3xF (UL33-3xF). Cells were next fixed and permeabilized over a 96 h time course, and UL33 was visualized via its FLAG epitopes (green). mCherry (red) is a marker of infection, and DAPI (blue) was used to visualize host cell nuclei. Images were acquired at 40X magnification. N = 3, representative images shown (B) Cells were infected as in (A) , with an additional, parallel culture treated with PAA post-adsorption, which was collected at 72 hpi. Cells were otherwise collected over a 96 h time course, and whole-cell lysates were immunoblotted for the FLAG epitope tag to detect pUL33-3xF. IE1 is shown as a marker of lytic infection, and actin is shown as a loading control. M, mock-infected cells, harvested at 96 hpi. N = 3, representative blots shown.

Article Snippet: Primary newborn human foreskin fibroblasts (NuFF-1) were obtained from GlobalStem.

Techniques: Infection, Marker, Adsorption, FLAG-tag, Control

Journal: bioRxiv

Article Title: The Human Cytomegalovirus vGPCR UL33 is Essential for Efficient Lytic Replication in Epithelial Cells

doi: 10.1101/2024.09.18.609710

Figure Lengend Snippet: Multistep growth analyses were performed in epithelial cells (ARPE-19 cells) infected (MOI= 0.1 TCID50/cell) with the indicated viruses over a 30-day time course. Infected cells were collected at the indicated times, and viral titers of cell-associated virus were quantified by TCID50 using naïve NuFF-1 fibroblasts. N = 3 biological replicates; graph depicts one biological replicate, with 3 technical replicates per time point. Error is shown as Standard deviation and significance was calculated using unpaired t-tests. * p <0.05, ** p <0.01, *** p <0.005

Article Snippet: Primary newborn human foreskin fibroblasts (NuFF-1) were obtained from GlobalStem.

Techniques: Infection, Virus, Standard Deviation

Journal: bioRxiv

Article Title: The Human Cytomegalovirus vGPCR UL33 is Essential for Efficient Lytic Replication in Epithelial Cells

doi: 10.1101/2024.09.18.609710

Figure Lengend Snippet: (A) Fibroblasts (NuFF-1) or (B) epithelial cells (ARPE-19) were mock-, WT-, or UL33Δ-infected (MOI= 0.5 TCID50/cell) for the times indicated. Whole cell lysates were prepared and representative viral proteins from the IE (IE1), E (pUL44), and L (pp28) kinetic classes were assessed by immunoblot. Actin was assayed as a loading control. N=3, representative blots shown.

Article Snippet: Primary newborn human foreskin fibroblasts (NuFF-1) were obtained from GlobalStem.

Techniques: Infection, Western Blot, Control