Journal: Advanced Healthcare Materials

Article Title: Generation of an Induced Pluripotent Stem Cell‐Derived Alveolar Type II In Vitro Model to Study Influenza A Virus Infection and Drug Treatments

doi: 10.1002/adhm.202405141

Figure Lengend Snippet: Differentiation, cultivation, and characterization of iAT2 cells. (a) Overview of the differentiation, cultivation, and plating procedure of iAT2s as described in the methods. iPSCs: induced pluripotent stem cells; iAT2s: iPSC‐derived alveolar type II cells; ALI: air–liquid interface; NKX2.1: NK2 homeobox 1; SP‐C: surface active agent protein C; for StemDiff, DS/SB, CBRa, CK/DCI, CK/DCI+Y: see methods. (b) Representative flow cytometry plots of the lung progenitor markers NKX2.1 coupled to GFP on day 14 (left) and SP‐C coupled to tdTomato on day 30 (right). (c) Representative transmission light image of alveolospheres within matrigel before passaging on day 35. Scale bar = 30 µm. (d) Representative transmission light images of iAT2s seeded on transwell inserts directly after seeding (day X) and 4 or 6 days after seeding (day X + 4 and X + 6). Scale bars = 50 µm. (e) Immunofluorescence images of iAT2s showing expression of antigen Kiel 67 (Ki67; green, left), SP‐C (magenta, left), lysophosphatidylcholine acyltransferase 1 (LPCAT1; yellow, middle), and NKX2.1 (green, right). Nuclei are displayed in blue. Single z‐planes are shown. Scale bars = 20 µm (left, middle) or 50 µm (right).

Article Snippet: Transwell inserts (Greiner Bio‐one, 662 641) were placed in 24‐well plates and coated with 100 μL of Corning Matrigel Human Embryonic Stem Cell‐qualified matrix (Corning, 354 277) according to the manufacturer's instructions.

Techniques: Derivative Assay, Flow Cytometry, Transmission Assay, Passaging, Immunofluorescence, Expressing

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