Journal: bioRxiv
Article Title: Membrane-Free Alveolus-on-a-Chip via Biodegradable Scaffold Recapitulates Interstitial Mechanics, Immune Trafficking, and Aerosolized mRNA Delivery
doi: 10.64898/2026.04.17.719302
Figure Lengend Snippet: (A) Schematic illustration of THP-1 monocyte migration across Transwell®, PCL, and PLGA membranes. Migration is restricted in Transwell® and PCL systems but enabled across the PLGA membrane. (B) Quantification of THP-1 cell migration after 24 h across epithelial–fibroblast co-cultures on the PLGA chip with or without MCP-1 (20 ng mL⁻¹), compared with Transwell® and PCL controls (n = 4). (C) Immunofluorescence images of THP-1 cells collected from the basolateral chamber after transmigrating through the fibroblast–epithelial layer. Cells were stained for CD14 (green) and F-actin (red), with nuclei counterstained with DAPI (blue). Scale bar: 50 μm. (D) Immunofluorescence images of THP-1 cells differentiated into macrophage-like cells following PMA treatment, stained for CD14 and F-actin (top) and CD68 and F-actin (bottom). Scale bar: 50 μm. (E) SEM image of macrophages on the chip under ALI conditions with fibroblast–epithelial co-culture. Scale bar: 30 μm. (F) Immunofluorescence images of lung fibroblasts, A549 cells, and THP-1–derived macrophages cultured on the alveoli-on-a-chip under ALI conditions. Fibroblasts were stained with TE-7, A549 cells were stained with ZO-1, macrophages were stained with CD68. The bottom panel shows a cross-sectional view highlighting the layered organization of the three cell types on the chip. Scale bars: 30 μm.
Article Snippet: THP-1 monocytes (ATCC, TIB-202) were differentiated into macrophage-like cells by treatment with phorbol 12-myristate 13-acetate (PMA, 50 ng mL−1) in RPMI 1640 medium for 24 h. After PMA stimulation, the medium was replaced with fresh PMA-free medium, and the cells were incubated for an additional 24 h to allow stabilization of the macrophage-like phenotype.
Techniques: Migration, Membrane, Immunofluorescence, Staining, Co-Culture Assay, Derivative Assay, Cell Culture
