Renewable Human Cell Model for Type 1 Diabetes Research: EndoC-βH5/HUVEC Coculture Spheroids

Area of Science:

• Biomedical Engineering
• Endocrinology
• Cell Biology

Background:

• In vitro drug screening for type 1 diabetes (T1D) relies heavily on human organ donor islets, which are scarce and heterogeneous.
• Limitations in human islet availability and donor variability hinder the development of standardized T1D therapies.
• There is a need for reproducible and scalable in vitro models to facilitate T1D drug discovery.

Purpose of the Study:

• To develop and evaluate a standardized 3D spheroid coculture model for in vitro type 1 diabetes research.
• To recapitulate islet morphology using immortalized beta-cells and human endothelial cells.
• To assess the functionality and viability of the 3D spheroid model for drug screening applications.

Main Methods:

• Coculture of immortalized human insulin-producing EndoC-βH5 cells with human umbilical vein endothelial cells (HUVECs) in 3D spheroids.
• Evaluation of different beta-cell/endothelial cell ratios (1:0, 1:1, 1:3) in spheroid formation.
• Assessment of insulin secretion, metabolic activity, cell viability, and gene expression.

Main Results:

• Compact spheroids (~140 μm diameter) formed within 7 days for monocultures and cocultures.
• The 1:3 EndoC-βH5/HUVEC spheroid ratio demonstrated significantly enhanced glucose-stimulated insulin secretion (over 20-fold increase) and ~12% higher metabolic activity compared to other groups.
• Treatment with glycine-modified INGAP-P further increased insulin stimulation in monoculture spheroids.

Conclusions:

• EndoC-βH5-based 3D spheroids provide a standardized and reproducible in vitro model for type 1 diabetes research.
• The 1:3 beta-cell/endothelial cell ratio enhances spheroid functionality, mimicking islet-like structures.
• This model shows promise for efficient in vitro drug screening and testing of T1D therapies.