human dermal fibroblasts df-f  (ZenBio)

Journal: bioRxiv

Article Title: Rapid Assessment of Biomarkers on Single Extracellular Vesicles Using ‘Catch and Display’ on Ultrathin Nanoporous Silicon Nitride Membranes

doi: 10.1101/2024.04.29.589900

Figure Lengend Snippet: (A-F) Representative regions of interest (ROI = 51 pm x 51 pm) in confocal microscope images taken following purified EV capture. EVs were collected from the conditioned cell culture media of mesenchymal stem cells (MSCs) and fibroblasts via ultracentrifugation and were evaluated using an antibody targeting CD9 and an appropriate isotype control (mouse [Ms] IgG). EVs were characterized with nanoparticle tracking analysis, labeled with carboxyfluorescein succinimidyl ester (CFSE) then antibodies, prefiltered with SepCon™ spin cups (0.5 pm pores), and diluted to prepare an input of 10 5 EVs. (A), (B), and (C) correspond with the CFSE signal, CD9 antibody signal, and merged signals (CFSE & CD9) fora representative MSC EV sample, respectively. (D), (E), and (F) correspond with the CFSE signal, CD9 antibody signal, and merged signals (CFSE & CD9) for a representative fibroblast EV sample, respectively. Arrows in (C) and (F) indicate instances of colocalization and detection of CD9+ EVs. Scale bars = 5 pm. (G) Detection summary for CD9 and isotype control in terms of % positive EVs for MSC and fibroblast samples. (H) CD9 detection summary for MSC and fibroblast EVs following isotype control correction. (I) Corresponding Western blot (WB) bulk assay comparison for CD9 expression in MSC and fibroblast EV samples. (J) Comparison of CD9 fold increase (MSC/fibroblast) as reported by CAD-LB and WB assays. All captured EVs were imaged with a fluorescence microscope and quantification and colocalization analyses were completed using an open-source FIJI plugin (ComDet v.0.5.5). All bars represent mean ± SEM (n = 3). * p < 0.05, *** p < 0.001, by Student’s t-test.

Article Snippet: Human dermal fibroblasts (ZenBio, Cat. # DF-F ) were cultured in low glucose Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific, Cat. # SH30021FS ) with 10% exosome-depleted FBS (Bio-Techne, Cat. # S11150 ; supernatant collected after centrifuging at 100,000⋅g for 18 hours) and 1% Antibiotic- Antimycotic (100X) (Thermo Fisher Scientific, Cat. # 15-240-062 ).

Techniques: Microscopy, Purification, Cell Culture, Control, Labeling, Western Blot, Comparison, Expressing, Fluorescence

Journal: Developmental cell

Article Title: Loss of Sirtuin 1 Alters the Secretome of Breast Cancer Cells by Impairing Lysosomal Integrity

doi: 10.1016/j.devcel.2019.03.011

Figure Lengend Snippet: KEY RESOURCE TABLE

Article Snippet: Human: Human Dermal Fibroblasts , Zenbio , DF-F.

Techniques: Recombinant, Isolation, Luciferase, Clone Assay, Activity Assay, shRNA, Sequencing, Plasmid Preparation, Software, SYBR Green Assay, Transfection

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: Comparison of TF, FVII, FIX, FX, and prothrombin protein levels measured in untreated ECs and fibroblasts. Cell lysates for quantification of ( a ) TF, ( b ) FVII, ( c ) FIX, ( d ) FX, and ( e ) prothrombin by ELISA were prepared from HUVECs (n = 3–8), LSECs (n = 3–4), GMVECs (n = 3–4), and fibroblasts (n = 3–5). Measured protein concentrations (means + SD) in pg/ml were normalized to total lysate protein to account for cell number difference. Values below the lowest detectable limit of the assay are noted as

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: Quantification of FVII, FIX, and FX released into the supernatants of untreated ECs and fibroblasts. HUVEC, LSEC, GMVEC, and fibroblast supernatants were collected from cells incubated for 24 hours in serum-free media and quantified by ELISA; ( a ) FVII, ( b ) FIX, and ( c ) FX. Graphs show protein concentrations in pg/ml (means + SD) for HUVECs (n = 4), GMVECs (n = 3–4), LSECs (n = 3–4), and fibroblasts (n = 3). *p < 0.05.

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: The effect of TNF and ATA exposure on HUVEC and fibroblast expression levels of THBD, PROCR , and TFPI , and HUVEC expression of F3, F9 , and F10 . The changes in gene expression of the surface coagulation regulatory proteins were measured in untreated HUVECs and fibroblasts (black), cells exposed to 10 ng/ml TNF (red), and cells exposed to 30 µg/ml ATA (blue) for 24 hours. Graphs show fold-changes in expression of ( a ) THBD, PROCR , and TFPI and ( b ) F3, F9 , and F10 in HUVECs and ( c ) THBD, PROCR , and TFPI in fibroblasts (means + SD). Number of gene measurements per experimental condition, HUVECs: THBD , n = 6; PROCR , n = 4–5; TFPI , n = 3; F3 , n = 3–4; F9 , n = 4; F10 , n = 3; for fibroblasts, each gene was measured 4 times under each experimental condition. *p < 0.05.

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: Expressing, Gene Expression, Coagulation

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: Comparison of FX activation on surfaces of HUVECs and fibroblasts. ( a ) FX activation was measured over time on HUVEC surfaces (in flasks and without addition of exogenous coagulation factors or phospholipids) by monitoring activated FX-mediated cleavage of the chromogenic substrate S-2765. The extent of FX activation was compared in untreated HUVECs (black, n = 11), HUVECs exposed to 10 ng/ml TNF (red, n = 6), and HUVECs exposed to 30 µg/ml ATA (blue, n = 7) for 24 hours. The FX activation in TNF-exposed HUVECs was significantly* higher than untreated HUVECs at time points 180 through 360 min; while ATA-exposed HUVECs generated significantly* less activated FX than untreated HUVECs at time points 240 to 360 min. ( b ) The extent of FX activation generated over time on surfaces of untreated HUVECs (black, n = 11) and fibroblasts (green, n = 4) is shown and compared with the chromogenic substrate S-2765 alone (blue, n = 6), demonstrating the absence of substrate self-hydrolysis. ( c ) FX activation was measured over time on fibroblast surfaces (in flasks and without addition of exogenous coagulation factors or phospholipids). The extent of FX activation was compared in untreated fibroblasts (black, n = 4), fibroblasts exposed to 10 ng/ml TNF (red, n = 4), and fibroblasts exposed to 30 µg/ml ATA (blue, n = 4) in serum-free media for 24 hours. *p < 0.05.

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: Comparison, Activation Assay, Coagulation, Generated

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: FX activation on fibroblast surfaces following the addition of human FVII and FX proteins. FX activation was measured on fibroblast surfaces by monitoring the hydrolysis of the chromogenic substrate S-2765 over 15 minutes. In a subset of measurements, plasma levels of either FVII (0.7 µg/ml), FX (10 µg/ml), or both were added to the calcium-containing buffer in the fibroblast flasks immediately before the addition of S-2765. There was minimal FX activation following addition of FVII alone (n = 5), increased FX activation following addition of FX alone (n = 5), and a marked increase in FX activation following the addition of both FVII and FX (n = 7) to the fibroblast containing-flasks.

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: Activation Assay, Clinical Proteomics

Journal: Scientific Reports

Article Title: Production and control of coagulation proteins for factor X activation in human endothelial cells and fibroblasts

doi: 10.1038/s41598-020-59058-4

Figure Lengend Snippet: Estimated levels of activated FX on HUVEC and fibroblast surfaces.

Article Snippet: Single donor human dermal fibroblasts were purchased from Zenbio (DF-F) at passage 2.

Techniques: