Journal: Materials Today Bio
Article Title: Synergistic aligned neuronal and vascular growth inside 3D-PEG-Anisogels utilizing a triple-co-culture
doi: 10.1016/j.mtbio.2025.102737
Figure Lengend Snippet: Effect of alignment on the triculture with single neurons. A, B: Respective images for immunofluorescence staining for NF-H (red) of single neurons (50/μL) and for PECAM/CD-31 (green) of structures formed by HUVECs (2000 cells/μL, ratio 3 MSCs: 1 HUVEC) in a control hydrogel (A) with their corresponding 40x image (B). C: Respective radial plots for the depicted control images' directionality of neurites and vascular-like structures. D,E: Respective images for immunofluorescence staining for NF-H (red) of single neurons (50/μL) and for PECAM/CD-31 (green) of structures formed by HUVECs (2000 cells/μL, ratio 3 MSCs: 1 HUVEC) in a a Anisogel with 1.00 v/v% 2.5x2.5 × 50 μm 3 microgels labeled with Rhodamine-B-acrylate (yellow) (D) and with their corresponding 40x image (E) with 2.00 w/v% PEG and 200 μM IKVAV; F: Respective radial plots for the depicted control images' directionality of neurites, vascular-like structures and microgels. G: 3D Analysis of the total length of vascular-like structures. H: 3D Analysis of the total neurite length from DRGs The PEG-QK gels were processed and analyzed after culturing for 7 days in EGM-2 + NGF. Scale bar: 1 mm (A, D); 200 μm (B, E). n: 3–11 replicates per condition (p∗ < 0.05, p∗∗ < 0.01, p∗∗∗ < 0.001). Error bars are given as standard error. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Article Snippet: Human umbilical vein endothelial cells (HUVECs, passage 1–5, pooled donors, Lonza) were cultured in tissue culture flasks in endothelial growth medium (EGM-2 ready-to-use kit, Promocell) supplemented with FBS (2.00 w/v%), epidermal growth factor (recombinant human, 5 ng/mL), basic fibroblast growth factor (recombinant human, 10.0 ng/mL), insulin-like growth factor (Long R3 IGF, 20 ng/mL), vascular endothelial growth factor 165 (recombinant human, 0.5 ng/mL), ascorbic acid (1 μg/mL), heparin (22.5 μg/mL), and hydrocortisone (0.20 μg/mL).
Techniques: Immunofluorescence, Staining, Control, Labeling