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