Journal: bioRxiv

Article Title: Modulating Hydrogel Stiffness Through Light-Based 3D Printing to Mimic Cardiac Fibrosis and Cardiomyocyte Dysfunction Using hiPSC-Derived Cells

doi: 10.1101/2025.05.20.655137

Figure Lengend Snippet: A) Day 20 HCFs were replated onto 3D printed hydrogel disks fabricated using 20 mW/cm 2 (left), 50 mW/cm 2 (middle), or 70 mW/cm 2 (right) light intensities, cultured for 48 hours in FGM-3 with 0.1 ng/mL TGFβ, and then stained for vimentin and αSMA. B) qPCR analysis comparing expression of genes associated with an activated cardiac fibroblast phenotype between quiescent HCFs (controls), HCFs cultured on tissue culture plates treated with TGFβ and HCFs cultured on hydrogels treated with or without TGFβ. Gene expression foldchange was calculated using the 2 -ΔΔCt method. C) Flow cytometry quantification of %αSMA+ in the same conditions as in panel B. D) Stratification of αSMA+ HCFs cultured on hydrogels and treated with TGFβ into “low” and “high” expression groups, based on fluorophore signal intensity, corresponding to lower and higher activation states, respectively. N=3 for each condition; *, **, ***, and **** indicate p-values ≤ 0.05, ≤ 0.01, ≤ 0.001, and ≤ 0.0001, respectively.

Article Snippet: The effects of activated HCFs on cardiac functionality were assessed via the in direct coculture of HCFs and hiPSC-derived cardiomyocytes (hiPSC-CMs) in 6-well, flat bottom tissue culture plates (Genesee Scientific, 25-105). hiPSC-CMs were generated using a small molecule-based approach for inhibiting Gsk3 and Wnt, as described by Lian et al. in 2013.

Techniques: Cell Culture, Staining, Expressing, Gene Expression, Flow Cytometry, Activation Assay