Journal: Journal of Biological Engineering

Article Title: Anti-CD31 antibody preconditioning for enhancement of endothelial cell capture and vascularization: a novel strategy for bioengineering lung scaffolds

doi: 10.1186/s13036-025-00593-x

Figure Lengend Snippet: Effect of coating agents on the attachment and growth of Endothelial cells (ECs) on precision cut lung slices (PCLS). ( A ) Schematic diagram describes the experimental design of each assay using decellularized PCLS. ( B , C ) Coating of decellularized scaffolds enhance the attachment of Hoechst 33,342-labeled (blue) mC166 cells on PCLS compared to the uncoated group. Each group n = 6. ( D ) Quantification of EC cell metabolic activity using cell growth assay using Cell Counting Kit-8 (CCK-8) revealed that precoating of PCLS increases ECs growth and metabolic activity (Optical Density (OD) measured at Day 1, 3 and 5) compared to ECs cultured on uncoated PCLS. Each group n = 6. ( E ) Quantification of the attachment of Hoechst-labeled HUVECs on anti-CD31 coated PCLS coated with 12.5, 25, 50, or 100 µg/mL anti-CD31 Ab. Results represent the mean of measurements taken from 3 PCLS seeded with HUVEC cells from 3 independent experiments. Each group n = 3. ( F ) Quantification of HUVEC cell metabolic activity using cell growth assay using Cell Counting Kit-8 (CCK-8) revealed that precoating of PCLS increases ECs growth and metabolic activity (Optical Density (OD) measured at Day 1, 3 and 5) compared to ECs cultured on uncoated PCLS. Each group n = 3. Scale bar = 200 μm. The results represent the mean of measurements taken from 3 technical replicates. Each number represents a biological replicate (independent experiment). M ean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Endothelial cell growth medium (EGM) (Lonza Bioscience, MD, USA) was used to culture human umbilical vein endothelial cells (HUVEC) (ATCC, Manassas, VA).

Techniques: Labeling, Activity Assay, Growth Assay, Cell Counting, CCK-8 Assay, Cell Culture