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Updated: May 8, 2026

Micropatterning and Assembly of 3D Microvessels

Micropatterning and Assembly of 3D Microvessels

Published on: September 9, 2016

A Simple Model to Study Mosaic Gene Expression in 3D Endothelial Spheroids.

Lucinda S McRobb¹, Vivienne S Lee¹, Fahimeh Faqihi¹

¹Macquarie Medical School, Faculty of Medicine, Health, and Human Sciences, Macquarie University, Sydney, NSW 2109, Australia.

Journal of Cardiovascular Development and Disease

|October 25, 2024

Summary

Researchers developed a simple 3D endothelial spheroid model using adeno-associated virus (AAV) for studying vascular malformations. This model enables mosaic gene expression and future analysis of post-zygotic mutations.

Keywords:

adeno-associated virus endothelial cells spheroid vascular malformations

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Area of Science:

Cell Biology
Vascular Biology
Biotechnology

Background:

3D cell culture models are crucial for studying complex biological processes.
Endothelial cells form blood vessels and are key in vascular development and disease.
Modeling mosaic mutations in endothelial cells is challenging but important for understanding vascular malformations.

Purpose of the Study:

To create a simple 3D endothelial spheroid model with mosaic gene expression.
To utilize adeno-associated virus (AAV) transduction for gene delivery.
To establish a platform for studying post-zygotic mutations in vascular malformations.

Main Methods:

Two immortalized human endothelial cell lines were cultured in 96-well plates.
Spontaneous aggregation and spheroid formation were monitored.
Adeno-associated virus (AAV) transduction was used to achieve mosaic gene expression (eGFP).

Main Results:

hCMEC/D3 cells spontaneously formed robust, stable 3D spheroids within 3 days.
These spheroids developed a basement membrane and expressed CD31.
AAV1 and AAV2^QUADYF transduction resulted in sustained, mosaic eGFP expression without toxicity.

Conclusions:

A simple method for generating 3D endothelial spheroids with mosaic gene expression was established.
This model facilitates the study of vascular malformations caused by mosaic mutations.
The model is suitable for assessing 3D phenotypes and drug/radiation sensitivities.