Abstract
The colony formation assay (CFA) is a widely used method to assess the self-renewal capacity of cancer cells and evaluate how this property is affected by drug treatment. This protocol presents a streamlined, high-throughput CFA workflow optimized for T-cell acute lymphoblastic leukemia (T-ALL), making it suitable for large-scale drug screening projects. Colonies are grown in a methylcellulose-based 3D matrix and quantified using an automated analysis pipeline, allowing robust estimation of colony number and size. This cost-effective approach provides a scalable platform for identifying compounds that impair self-renewal, facilitating prioritization of hits for validation in more complex in vivo models.
Document Type
Article
Publication Date
2025
Digital Object Identifier (DOI)
https://doi.org/10.1016/j.scr.2025.103757
Funding Information
This research was supported by the National Cancer Institute (R37CA227656 to JSB) and the Kentucky Pediatric Cancer Research Trust Fund (PON2 728 2400001524 to JSB).
Repository Citation
Al-Hamaly, Majd and Blackburn, Jessica S., "A high-throughput workflow for assessing self-renewal using colony formation assays" (2025). Markey Cancer Center Faculty Publications. 469.
https://uknowledge.uky.edu/markey_facpub/469
Notes/Citation Information
1873-5061/© 2025 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).