Additional file 1 of Anti-CD31 antibody preconditioning for enhancement of endothelial cell capture and vascularization: a novel strategy for bioengineering lung scaffolds

Supplementary Material 1: Fig: 1 Decellularization of the mouse lungs. (A) Gross appearance of native lung and decellularized lung demonstrates the whitish coloration and preservation of the shape and size of the scaffold after the decellularization process. (B) Histological appearance after H&E staining of decellularized lung showing no cellular material compared to native lung. Scale bar = 100 μm. (C) Confocal microscopy images showing nuclear DAPI staining of native and decellularized lung tissues reveals total removal of nuclei from decellularized lung tissues. (D) DNA quantification of native lung and decellularized scaffolds indicating that only a negligible amount of DNA is retained compared to that in native lung. The results represent the mean of triplicate measurements. Each number (n=) represents a biological replicate (independent decellularized lung scaffold). Each group n = 6, mean ± SD, **p < 0.01. (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. (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. The results represent the mean of measurements taken from 3 technical replicates. Each number (n=) represents a biological replicate (independent experiment). Each group n = 3, mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.