Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Morphological and immunocytochemical fibroblast identification. Representative brightfield and immunofluorescence images of the fibroblast markers vimentin, DDR2, collagen 1, and αSMA. The nuclei were stained with DAPI (blue). Upper panel) HVFs. Mid panel) HAFs. Lower panel) PAFs. The scale bars equal 50 µm.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Immunofluorescence, Staining

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Induction of a cellular fibrosis phenotype in HAFs with TGF‐β. (A) Protein expression of phosphorylated SMAD2/3 after 72 h of TGF‐β stimulation (1, 3, 10 ng·mL −1 ) in HAFs ( n = 3 per concentration) and representative original WB below. (B) Protein expression of αSMA after 72 h of TGF‐β stimulation (1, 3, 10 ng·mL −1 ) in HAFs ( n = 3 per concentration) and representative original WB below. (C) Quantification of fibroblasts positive for fibrillary αSMA microfilaments after stimulation with 10 ng·mL −1 TGF‐β for 72 h and representative immunofluorescence staining of fibrillary αSMA upon TGF‐β stimulation. The scale bars equal 50 µm. (D) Soluble collagen secretion (left) and representative immunofluorescence image for deposited Collagen Iα1 (right) by HAFs upon stimulation with 10 ng·mL −1 TGF‐β ( n = 14 vs. 10). The scale bars equal 50 µm. (E) Proliferation curves of HAFs under control conditions ( n = 4) and upon stimulation with 10 ng·mL −1 TGF‐β ( n = 4). Cells were counted after 7 and 14 days. (F) 24‐h migration capacity of HAFs under control conditions ( n = 6) and upon stimulation with 10 ng·mL −1 TGF‐β ( n = 4). Data are presented as mean ± SEM. Differences between two groups were compared using Student’s t ‐test with Welch’s correction. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. * P < 0.05. ** P < 0.01. *** P < 0.001.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Expressing, Concentration Assay, Immunofluorescence, Staining, Control, Migration

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Functional analysis of cardiac fibroblast subtypes. (A) Proliferation curves of HVFs ( n = 6), HAFs ( n = 7) and PAFs ( n = 21). Cells were cultured in DMEM (10% FCS, 1% penicillin–streptomycin) at 37 °C, 5% CO 2 , and counted after 7 and 14 days. (B) Quantification of immunostaining experiments for fibrillary αSMA protein abundance ( n HVF = 4, n HAF = 7, n PAF = 6). (C) Basal 24‐h migration capacity of HVFs ( n = 5), HAFs ( n = 5), and PAFs ( n = 6). Data are presented as mean ± SEM. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. * P < 0.05. ** P < 0.01. *** P < 0.001; n.s., not significant.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Functional Assay, Cell Culture, Immunostaining, Quantitative Proteomics, Migration

Journal: FEBS Open Bio

Article Title: Modeling atrial fibrosis in vitro —Generation and characterization of a novel human atrial fibroblast cell line

doi: 10.1002/2211-5463.12896

Figure Lengend Snippet: Adaption of HAF and PAF stiffness in response to different stiffness of the growth matrix. (A) HAFs present a typical fibroblast morphology when grown on CyPhyGels. Nuclei were stained with Hoechst (blue), F‐actin was stained with Phalloidin (red), and ɑSMA was stained in green. The scale bar equals 20 µm. (B) Representative force/ indentation curves used to calculate the stiffness (E eff ) of individual cells cultured on either stiff (black curve) or soft gels (grey curve). The force required to indent a cell on the stiff substrate is higher than on the soft substrate. (C) Measurements of HAF and PAF stiffness on soft (~2.7 kPa) and stiff (~4.6 kPa) CyPhyGels (36 ≤ n ≤ 57). Data are presented as mean ± SEM. Differences between multiple groups were compared with one‐way ANOVA with Newman–Keuls post‐test. *** P < 0.001.

Article Snippet: Using lentiviral transfer of Upcyte ® proliferation genes, a novel human atrial fibroblast cell line (HAF‐SRK01) was generated.

Techniques: Staining, Cell Culture