hcf media  (PromoCell)

Journal: iScience

Article Title: Cardiomyocyte-fibroblast interaction regulates ferroptosis and fibrosis after myocardial injury

doi: 10.1016/j.isci.2024.109219

Figure Lengend Snippet: Fibroblast-derived cytokine and chemokine promote iCM survival after erastin treatment (A) Flowchart of conditioned media and cytokine array experiment. (B) Survival rate of iCMs, cultured in control medium (DMEM), HCF conditioned medium, or H 2 O 2 -treated HCF conditioned medium, after erastin treatment, normalized to respective DMSO control groups. (C) Cytokine-chemokine protein array blotting image of conditioned media from vehicle- or H 2 O 2 -treated HCFs. (D) Blotting signal intensity of IL-8 and EGF. (E and F) Survival rate of iCMs after erastin treatment in the presence of IL-8 (E) or EGF (F), normalized to DMSO control groups. (G) qPCR of CXCR1 and CXCR2 in total blood cells and purified cardiomyocyte nuclei after P1 LAD-O. (H) Western blot of CXCR1, CXCR2, and α-tubulin in human iCMs after DMSO or erastin treatment. (I) Normalized band (immuno-blotting) intensity of CXCR1 and CXCR2 in (H). All bar graphs represent mean ± SD. ∗p < 0.05; ∗∗p < 0.01; NS, not significant by t test.

Article Snippet: Human Cardiac Fibroblasts (HCF) (Cell Applications Inc, 306V-05a) were cultured in HCF Growth Medium (Cell Applications Inc, 316–500).

Techniques: Derivative Assay, Cell Culture, Control, Protein Array, Purification, Western Blot

Journal: iScience

Article Title: Cardiomyocyte-fibroblast interaction regulates ferroptosis and fibrosis after myocardial injury

doi: 10.1016/j.isci.2024.109219

Figure Lengend Snippet: Cardiac fibroblasts interact with cardiomyocytes through gap junctions to share free iron (A and B) Wild-type mouse heart tissue stained for Fth1 [magenta, (A)] or Ftl [magenta, (B)], with cTnT (green) and DAPI (blue) at 1DPMI after P7 LAD-O. Arrows: non-cardiomyocytes positive for Fth1 (A) or Ftl (B). (C and D) Mouse heart tissue stained for Fth1 [red, (C)] or Ftl [red, (D)], with Pdgfrα (gray), cTnT (green), and DAPI (blue) at 1 DPMI after P7 LAD-O. Arrows: cells positive for Pdgfrα and Fth1 (C) or Ftl (D). (E and F) Mouse heart tissue stained for Fth1 [green, (E)] or Ftl [green, (F)], with Pdgfrα (red), MF20 (gray), and DAPI (blue) at 6 DPMI after P7 LAD-O. Arrows: cells positive for Pdgfrα and Fth1 (E) or Ftl (F). (G) Diagram of cardiomyocyte-fibroblast interaction after MI. (H and I) Mouse heart section stained for Cx45 [green, (H)] or Cx43 [green, (I)], with Pdgfrα (red), MF20 (gray), and DAPI (blue) after P7 LAD-O. Arrows: potential locations of gap junctions between cardiomyocytes and fibroblasts. (J–L) Co-cultured iCM and HCF stained for VIMENTIN (VIM, gray), free Fe 2+ (red), DAPI (blue), and imaged with TITIN-GFP (green) after DMSO (J) or erastin (15 μM) (K, L) treatment. Asterisks: HCFs with accumulation of Fe 2+ . (M) siRNA knockdown of CX43 and CX45 simultaneously in iCM-HCF co-culture; Fe 2+ fluorescent intensity ratio of iCMs over HCFs was quantified after erastin or DMSO treatment. LV, left ventricle. All bar graphs represent mean ± SD. ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001 by t test. Scale bar, 75 μm (A, B, E, F, H), 25 μm (C, D, I, J–L). See also Figures S4 and .

Article Snippet: Human Cardiac Fibroblasts (HCF) (Cell Applications Inc, 306V-05a) were cultured in HCF Growth Medium (Cell Applications Inc, 316–500).

Techniques: Staining, Cell Culture, Knockdown, Co-Culture Assay

Journal: iScience

Article Title: Cardiomyocyte-fibroblast interaction regulates ferroptosis and fibrosis after myocardial injury

doi: 10.1016/j.isci.2024.109219

Figure Lengend Snippet:

Article Snippet: Human Cardiac Fibroblasts (HCF) (Cell Applications Inc, 306V-05a) were cultured in HCF Growth Medium (Cell Applications Inc, 316–500).

Techniques: Recombinant, Produced, Virus, Bicinchoninic Acid Protein Assay, Purification, Sequencing, Negative Control, Software

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Vesicles Secreted by Corneal Myofibroblasts Promote Corneal Epithelial Cell Migration

doi: 10.3390/ijms23063136

Figure Lengend Snippet: Isolation and characterization of human corneal keratocyte (HCK)-, fibroblast (HCF)-, and myofibroblast (HCM)-derived extracellular vesicles (EVs). ( A ) EV pellets (20 μg protein/lane) were analyzed by western blot to probe for vesicle-associated markers. Representative images are shown for vesicle-associated proteins CD63, CD81, and ITGAV; a negative control (GM130); and proteins associated with corneal wound healing, FN1 and THBS1. ( B ) EV pellets were analyzed by nanoparticle tracking analysis (NTA), and a size distribution histogram for each EV sample is shown. (( B ), inset) Transmission electron microscopy images demonstrating EV morphology (high magnification, 49000x; scale bar = 100 nm). ( C ) The average particle concentrations (particles x1011/mL); ( D ) mean particle size (nm); and ( E ) zeta (ζ) potential of EV pellets were measured using NTA with Zetaview™. Data are shown as mean + SEM; n = 3 independent EV preparations; ns = nonsignificant. One-way ANOVA with Tukey’s post-test.

Article Snippet: Once isolated, cells were plated on 6-well plates and grown to 75% confluency in complete Eagle’s Minimum Essential Media (EMEM (American Type Culture Collection; Manassas, VA, USA) with 1× antibiotic-antimycotic (Gibco)) and either 1% (HCK media) or 10% (HCF media) fetal bovine serum (FBS, Atlanta Biologicals; Flowery Branch, GA, USA).

Techniques: Isolation, Derivative Assay, Western Blot, Negative Control, Transmission Assay, Electron Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Vesicles Secreted by Corneal Myofibroblasts Promote Corneal Epithelial Cell Migration

doi: 10.3390/ijms23063136

Figure Lengend Snippet: Proteomic analysis of corneal keratocyte, fibroblast, and myofibroblast extracellular vesicles. ( A ) The heatmap of differentially expressed proteins from human corneal keratocyte (HCF) and fibroblast (HCF) relative to myofibroblast (HCM) extracellular vesicles. A multigroup comparison was performed and showed that 195 proteins were differentially expressed in our dataset; proteins selected show log-fold change values of target proteins (blue (−3.5) to yellow (+3.5) through white), ranked by adjusted p < 0.05. ( B ) The top 25 proteins are selected from this heatmap with decreased (blue (−3.5)) and increased (yellow (+3.5)) log-fold change in HCK-EVs and HCF-EVs relative to HCM-EVs. Data shown as n = 3 independent EV preparations.

Article Snippet: Once isolated, cells were plated on 6-well plates and grown to 75% confluency in complete Eagle’s Minimum Essential Media (EMEM (American Type Culture Collection; Manassas, VA, USA) with 1× antibiotic-antimycotic (Gibco)) and either 1% (HCK media) or 10% (HCF media) fetal bovine serum (FBS, Atlanta Biologicals; Flowery Branch, GA, USA).

Techniques: Comparison

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Vesicles Secreted by Corneal Myofibroblasts Promote Corneal Epithelial Cell Migration

doi: 10.3390/ijms23063136

Figure Lengend Snippet: Ingenuity pathway analysis of proteins from corneal myofibroblast extracellular vesicles. ( A ) The top 10 disease and biological functions identified from the ingenuity pathway analysis (IPA) using the dataset of 195 proteins. The p -value for each biological function is indicated by the bar and is expressed as -log 2 ( p -value). The orange dashed line indicates the threshold of significance ( p < 0.05). ( B ) The top-scoring proteins identified by IPA for cellular movement; green indicates decreased measurement, red indicates increased measurement, blue dashed arrow leads to inhibition. ( C ) The heatmap shows 31 proteins with decreased (blue (−3.5)) and increased (yellow (+3.5)) log-fold change in HCK-EVs and HCF-EVs relative to HCM-EVs. Data shown as n = 3 independent EV preparations.

Article Snippet: Once isolated, cells were plated on 6-well plates and grown to 75% confluency in complete Eagle’s Minimum Essential Media (EMEM (American Type Culture Collection; Manassas, VA, USA) with 1× antibiotic-antimycotic (Gibco)) and either 1% (HCK media) or 10% (HCF media) fetal bovine serum (FBS, Atlanta Biologicals; Flowery Branch, GA, USA).

Techniques: Inhibition