human cardiac fibroblasts cell culture  (Innoprot Inc)

Journal: Nature communications

Article Title: DNMT3A clonal hematopoiesis-driver mutations induce cardiac fibrosis by paracrine activation of fibroblasts.

doi: 10.1038/s41467-023-43003-w

Figure Lengend Snippet: Fig. 7 | CHIP monocyte-derived HB-EGF induces cardiac fibroblasts activation. a Relative DNMT3A expression and (b) HBEGF expression in DNMT3A-deficient PMA- activated THP-1 cells normalized to RPLP0 mRNA expression analyzed by qPCR (n = 12 biologically independent samples). Source data are provided as a Source Data file. c Relative DNMT3A and HB-EGF protein expression in DNMT3A silenced PMA- activated THP-1 cells (n = 4 biologically independent experiments). Source data are provided as a Source Data file. d Immunofluorescence analysis of αSMA protein expression in HB-EGF (100 ng/ml, 48 h) stimulated iHCF (n = 3 biologically inde- pendent experiments). Source data are provided as a Source Data file. e Immunofluorescence analysis images of phospho-histone H3 protein expression in HB-EGF stimulated HCF (n = 4 biologically independent experiments). Source data are provided as a Source Data file. f Beating frequency of cardiospheres stimulated with HB-EGF (100 ng/ml, 5 days) (n = 4 for controls and n = 6 for HB-EGF, biologically independent experiments). Source data are provided as a Source Data file.

Article Snippet: Human cardiac fibroblasts cell culture and treatment Primary human cardiac fibroblasts (HCF) (#C-12375, Promocell) or immortalized human cardiac fibroblasts (iHCF) (#P10453-IM, Innoprot) were used.

Techniques: Derivative Assay, Activation Assay, Expressing

Journal: Experimental Biology and Medicine

Article Title: Featured Article: TGF-β1 dominates extracellular matrix rigidity for inducing differentiation of human cardiac fibroblasts to myofibroblasts

doi: 10.1177/1535370218761628

Figure Lengend Snippet: Image processing techniques and experimental timeline. (a) Representative raw and thresholded images of nuclei, actin, and α-SMA in untreated and TGF-β1-treated human cardiac fibroblasts (scale bars: 200 μm). (b) Experimental timeline for culturing cardiac fibroblasts with reduced serum and TGF-β1 treatment. (A color version of this figure is available in the online journal.)

Article Snippet: Cell culture Primary human adult cardiac fibroblasts (Lot 3131, Cell Applications Inc., San Diego, CA, USA) were thawed into a 75 cm 2 cell culture flask with fibroblast growth medium, consisting of low glucose DMEM (1 g/L glucose) supplemented with 10% v/v fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 1% v/v penicillin-streptomycin solution (10,000 U/mL penicillin, 10,000 μg/mL streptomycin).

Techniques:

Journal: Experimental Biology and Medicine

Article Title: Featured Article: TGF-β1 dominates extracellular matrix rigidity for inducing differentiation of human cardiac fibroblasts to myofibroblasts

doi: 10.1177/1535370218761628

Figure Lengend Snippet: Cell morphology and α-SMA expression due to ECM rigidity and TGF-β1 treatment. Representative images of human cardiac fibroblasts cultured for five days on low, moderate, and high PDMS-coated coverslips with or without 2 ng/mL TGF-β1 (blue: nuclei, green: actin, red: α-SMA, scale bars: 25 μm).

Article Snippet: Cell culture Primary human adult cardiac fibroblasts (Lot 3131, Cell Applications Inc., San Diego, CA, USA) were thawed into a 75 cm 2 cell culture flask with fibroblast growth medium, consisting of low glucose DMEM (1 g/L glucose) supplemented with 10% v/v fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 1% v/v penicillin-streptomycin solution (10,000 U/mL penicillin, 10,000 μg/mL streptomycin).

Techniques: Expressing, Cell Culture

Journal: Experimental Biology and Medicine

Article Title: Featured Article: TGF-β1 dominates extracellular matrix rigidity for inducing differentiation of human cardiac fibroblasts to myofibroblasts

doi: 10.1177/1535370218761628

Figure Lengend Snippet: Quantification of cell morphology and α-SMA expression due to ECM rigidity and TGF-β1 treatment. Human cardiac fibroblasts were cultured on low, moderate, and high PDMS-coated coated coverslips with or without 2 ng/mL TGF-β1 for one, three, and five days. Cells were immunostained and cell density (a), actin coverage (b), and α-SMA/actin coverage (c) were quantified from immunostained images (n = 4, bars indicate mean ± standard error of the mean, *P < 0.05 and **P < 0.01 according to one-way ANOVA and Tukey’s test for multiple comparisons. For detailed statistical analysis, see Supplementary Tables S2 to S4). (A color version of this figure is available in the online journal.)

Article Snippet: Cell culture Primary human adult cardiac fibroblasts (Lot 3131, Cell Applications Inc., San Diego, CA, USA) were thawed into a 75 cm 2 cell culture flask with fibroblast growth medium, consisting of low glucose DMEM (1 g/L glucose) supplemented with 10% v/v fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 1% v/v penicillin-streptomycin solution (10,000 U/mL penicillin, 10,000 μg/mL streptomycin).

Techniques: Expressing, Cell Culture

Journal: Experimental Biology and Medicine

Article Title: Featured Article: TGF-β1 dominates extracellular matrix rigidity for inducing differentiation of human cardiac fibroblasts to myofibroblasts

doi: 10.1177/1535370218761628

Figure Lengend Snippet: Relative changes in gene expression due to ECM rigidity and TGF-β1 treatment. Human cardiac fibroblasts were cultured on low, moderate, and high PDMS-coated coated coverslips with or without 2 ng/mL TGF-β1 for one, three, and five days. RT-PCR was used to quantify the relative expression of (a) ACTA2, (b) POSTN, (c) FAP, (d) FSP1, and (e) GJA1. All data were normalized to high PDMS control group on Day 1. (n = 4, bars indicate mean ±standard error of the mean, *P < 0.05 and **P < 0.01 according to one-way ANOVA and Tukey’s test for multiple comparisons. For detailed statistical analysis, see Supplementary Tables S5 to S9). (A color version of this figure is available in the online journal.)

Article Snippet: Cell culture Primary human adult cardiac fibroblasts (Lot 3131, Cell Applications Inc., San Diego, CA, USA) were thawed into a 75 cm 2 cell culture flask with fibroblast growth medium, consisting of low glucose DMEM (1 g/L glucose) supplemented with 10% v/v fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 1% v/v penicillin-streptomycin solution (10,000 U/mL penicillin, 10,000 μg/mL streptomycin).

Techniques: Gene Expression, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Expressing, Control