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: Infection and Immunity

Article Title: Differential sensitivity of leukocyte populations to Staphylococcus aureus biofilm

doi: 10.1128/iai.00654-25

Figure Lengend Snippet: Direct contact with S. aureus biofilm is critical to induce leukocyte cytotoxicity. Primary Mφs, G-MDSCs, and PMNs were co-cultured with S. aureus biofilm either directly or separated by Transwell inserts for the indicated intervals. Leukocytes were stained with anti-CD45, Zombie NIR (viability), Apotracker Green (phosphatidylserine), and MitoSOX Red (mtROS) to quantify ( A ) leukocyte viability, ( B ) early apoptosis, ( C ) late apoptosis/necrosis, and mtROS levels in ( D ) live and ( E ) early apoptotic cells. Unstimulated leukocytes were incubated in medium for 30 min or 2 h, as indicated ( n = 9 from three independent experiments; *, P < 0.05; **, P < 0.01; ****, P < 0.0001; one-way ANOVA with Dunnett’s multiple correction between cell types).

Article Snippet: On day 4 of biofilm growth, ~45% of the medium was removed and leukocytes were added at a density of 2.5 × 10 5 cells/well in fresh medium and incubated for 15 min, 30 min, 2 h, and 6 h. For Transwell co-cultures, Transwell inserts (0.4 μm; CELLTREAT #230635) were placed in 24-well plates above the biofilm on day 4 of growth, whereupon 7.5 × 10 5 leukocytes were added to the upper chamber for 30 min or 2 h. Leukocytes were stained and acquired as described above for planktonic co-culture studies, and unstimulated cells were included for the duration of the co-culture period.

Techniques: Cell Culture, Staining, Incubation