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Related Experiment Video

Updated: May 8, 2025

Three-Dimensional 3D Tumor Spheroid Invasion Assay

Three-Dimensional 3D Tumor Spheroid Invasion Assay

Published on: May 1, 2015

Towards a more objective and high-throughput spheroid invasion assay quantification method.

Rozanne W Mungai¹, Roger J Hartman², Grace E Jolin¹

¹Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, MA, 01605, USA.

Scientific Reports

|December 28, 2024

Summary

This summary is machine-generated.

A new high-throughput method objectively quantifies 3D cell invasion from multicellular spheroids in hydrogels. This approach minimizes spheroid size variations and provides precise, directionally-dependent invasion metrics for diverse biological applications.

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Last Updated: May 8, 2025

Three-Dimensional 3D Tumor Spheroid Invasion Assay

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Published on: May 1, 2015

Assessment of Ovarian Cancer Spheroid Attachment and Invasion of Mesothelial Cells in Real Time

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Published on: August 8, 2019

Area of Science:

Biomedical Engineering
Cell Biology
Quantitative Biology

Background:

Multicellular spheroids in 3D hydrogels are key in vitro models for studying cell invasion.
Existing quantification methods for spheroid cell invasion lack high-throughput, objectivity, and accessibility.
Variations in spheroid size and cell shape complicate objective assessment of invasion extent.

Purpose of the Study:

To develop a high-throughput, objective quantification method for 3D cell invasion from spheroids.
To minimize sensitivity to initial spheroid size and cell spreading.
To provide precise, directionally-dependent invasion metrics.

Main Methods:

Analysis of fluorescent cell nuclei images to automatically calculate invasion metrics at the pixel level.
Segmentation of the initial spheroid boundary and automated calculation of nuclear pixel distances.
Introduction of the area moment of inertia as an integrative invasion metric and principal component analysis for directional stimuli influence.

Main Results:

The developed method enables automated, pixel-level calculation of invasion metrics.
New metrics, including area moment of inertia, provide integrative and directionally-dependent invasion assessments.
The method successfully analyzed the invasiveness of five different cell types, demonstrating its power and utility.

Conclusions:

The high-throughput quantification method provides consistent and objective analysis of 3D multicellular spheroid invasion.
The method is adaptable for various cell types and biological applications like cancer metastasis and wound healing.
Analysis code in MATLAB and Python, along with a GUI, is provided for broad accessibility.