A high-throughput workflow for assessing self-renewal using colony formation assays

Majd A. Al-Hamaly; Jessica S. Blackburn

doi:10.1016/j.scr.2025.103757

A high-throughput workflow for assessing self-renewal using colony formation assays

Majd A. Al-Hamaly
, Jessica S. Blackburn

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

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.

Original language	English
Article number	103757
Journal	Stem Cell Research
Volume	87
DOIs	https://doi.org/10.1016/j.scr.2025.103757
State	Published - Sep 2025

Bibliographical note

Publisher Copyright:
© 2025 The Authors

Funding

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).

Funders	Funder number
National Childhood Cancer Registry – National Cancer Institute	R37CA227656
Kentucky Pediatric Cancer Research Trust Fund	PON2 728 2400001524

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Colony formation assay
Drug screening
High-throughput
Leukemia
Self-renewal

ASJC Scopus subject areas

General Medicine
Developmental Biology
Cell Biology

Access to Document

10.1016/j.scr.2025.103757

Cite this

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title = "A high-throughput workflow for assessing self-renewal using colony formation assays",

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.",

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language = "English",

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AU - Blackburn, Jessica S.

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AB - 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.

KW - Colony formation assay

KW - Drug screening

KW - High-throughput

KW - Leukemia

KW - Self-renewal

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M3 - Article

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AN - SCOPUS:105008878757

SN - 1873-5061

VL - 87

JO - Stem Cell Research

JF - Stem Cell Research

M1 - 103757

ER -

A high-throughput workflow for assessing self-renewal using colony formation assays

Abstract

Bibliographical note

Funding

UN SDGs

Keywords

ASJC Scopus subject areas

Access to Document

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