Journal: Stem Cell Reports
Article Title: Aged human serum induces vascular changes in an isogenic co-culture venule model
doi: 10.1016/j.stemcr.2025.102544
Figure Lengend Snippet: Permeability and transport of low-density lipoprotein and transferrin (A) Representative images of pHrodo Green-low-density lipoprotein (LDL) permeability in co-culture venule-like models under perfusion with young or aged serum with or without methyl-beta-cyclodextrin (MβCD), a multi-pathway endocytosis inhibitor. (B) Quantitative permeability of LDL before serum perfusion (day 0) and after 4 days of young (+YS) or aged serum (+AS) perfusion in co-culture venule-like models (day 4), n = 4 independent microvessels and experiments for young serum perfusion at day 4, n = 3 for all other conditions. (C) Quantitative permeability of LDL after 4 days of perfusion with YS with or without pre-treatment with MβCD. n = 3 independent microvessels and experiments for all conditions. (D) Quantitative permeability of LDL after 4 days of perfusion with AS with or without MβCD; chloroquine, a clathrin-dependent endocytosis inhibitor; or 7-ketocholesterol, a clathrin-independent endocytosis inhibitor. n = 3 independent microvessels and experiments for all conditions. (E) Representative images of transferrin (Alexa Fluor 647) permeability in co-culture venule-like models under perfusion with young or aged serum with or without chloroquine. (F) Quantitative permeability of transferrin after 4 days perfusion with young or aged serum in co-culture venule-like models. n = 3 independent microvessels and experiments for all conditions. (G) Quantitative permeability of transferrin after 4 days perfusion with young serum in co-culture venule-like models with or without chloroquine. n = 3 independent microvessels and experiments for all conditions. (H) Quantitative permeability of transferrin after 4 days perfusion with aged serum in co-culture venule-like models with or without chloroquine. n = 3 independent microvessels and experiments for all conditions.
Article Snippet: To assess barrier permeability, microvessels were perfused with different molecular weight fluorescently-labeled solutes after 4 days of perfusion with the appropriate serum condition: 200 μM LY (Sigma, CH dilithium salt) and 2 μM Alexa Fluor 647-conjugated 10 kDa dextran (Thermo Fisher Scientific, cat. no. D22914 ), 2 μM Texas Red-conjugated 70 kDa dextran (Thermo Fisher Scientific, D1864) and 2 μM fluorescein-conjugated 2 MDa dextran (Thermo Fisher Scientific, D7137), 100 μg mL −1 pHrodo Green LDL (Thermo Fisher Scientific, L34355 ), or 100 μg mL −1 transferrin from human serum conjugated to Alexa Fluor 647 (Thermo Fisher Scientific, T23366 ) in iEC media.
Techniques: Permeability, Co-Culture Assay
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