Authors:

Ethan Lee

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Organoids are revolutionary miniature human tissue models made from pluripotent stem cells that replicate the native tissue structures and functions of human organs. However, organoids that lack a vascular system for gas and nutritional exchange will eventually experience necrosis or poor differentiation in the interior as they grow larger in size [1]. To address this diffusion limit, this project focuses on engineering vascularized spinal cord organoids (SCOs) by integrating human umbilical vein endothelial cells (HUVECs) using bioactive peptide amphiphiles (PAs). In this study, preliminary trials were conducted to determine the optimal concentration and type of bioactive PAs, designed to promote vascular growth (VEGF-A PA) and cell proliferation (IKVAV PA) [2]. Histology markers for proliferation, morphology, and phosphorylation of VEGF-A receptors (VEGF-A signaling) determined that the ideal treatment conditions consist of micelle-forming VEGF-A PAs and fiber-forming IKVAV PAs. Following these trials, a 3D co-culture system was established by introducing HUVECs to SCOs. To optimize endothelial cell integration and penetration into the organoid core, three distinct treatment timelines were examined: pre-treating SCOs with PAs two days before HUVEC introduction, simultaneous application of PAs and HUVECs, and delayed PA treatment following initial co-culture. Co-cultures were maintained between 3 to 14 days to monitor the rate of cellular integration and endogenous proliferation. Results will be evaluated through immunostaining and imaging to analyze HUVEC penetration and VEGF-A phosphorylation (signaling) within the SCOs. By leveraging the potent bioactivity of PAs, this project aims to use vascular engineering to develop in vitro models that more accurately replicate human tissue. These improved models will facilitate future studies of spinal cord injuries and the development of regenerative therapeutic strategies.

Source:

Northwestern University

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Co-authors:

Ethan Lee