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Tissue Engineering

Tissue Engineering - Poster Session B

Poster K20 - Development of 3D Tissue Models for Microvasculature Studies

Thursday, October 24, 2024
2:45 PM – 3:45 PM EST
Location: Exhibit Hall E, F & G

Introduction:  Development of vascularized tissue is one of the key challenges for the successful clinical application of cell therapies, tissue engineered constructs, and other cardiac therapeutic approaches. Despite the significant efforts, establishing vascularized tissues remains a significant challenge, particularly in pathological tissue conditions. Engineering organized vascular networks is a crucial step in the development of functional and clinically relevant 3D diseased tissue models. Changes in the composition of the ECM microenvironment during tissue remodeling may alter angiogenic signaling and endothelial responsiveness. The goal of this project is to develop 3D tissue models that facilitate the study/discovery of molecules of therapeutics that can improve vascularization during tissue repair and regeneration. The objective of this study is to investigate how ECM composition influences endothelial microvasculature formation in a 3D vitro model.

Materials and

Methods:  Human umbilical vein endothelial cells (HUVECs) were seeded (45,000 cells/cm2) on fibrin gels and on Collagen Type I hydrogels at two final concentrations, 2.0 mg/mL and 4.0 mg/mL. HUVECs were also co-cultured with fibroblast cells at a ratio of 30:1 (HUVEC:Fibroblast). Samples were cultured for 24h, 48h, or 72h. The samples were then fixed and stained using Alexa-Phalloidin for F-actin, and DAPI for the nuclei.

Results, Conclusions, and Discussions: Preliminary imaging results indicate that HUVECs co-cultured with fibroblasts show improved in vitro microvasculature development when compared with HUVEC cultures without fibroblasts. Fibrin gels supported microvasculature development for the short-term cultures (24h and 48h), but at 72h microvasculature seems to have decreased. Collagen gels (2.0 mg/mL) were able maintain some microvasculature formation for 24h, 48h, and 72h. Lower density collagen gels (2.0mg/mL) seem to better support microvasculature formation compared to the higher concentration collagen gels (4.0 mg/mL). Overall, current results indicate that extracellular matrix composition strongly influences micro-vessel formation, and that maintenance of robust vessels is influenced by other cells in the microenvironment. Current work is focused on optimizing the development of these 3D models and further characterizing microvasculature formation in healthy and in fibrotic conditions in vitro. These engineered vascularized models could enable the evaluation of novel therapeutics.

Acknowledgements (Optional):  This research was supported by the National Science Foundation CAREER Award, as well as the NSF REU at CSULB.