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Related Experiment Video

Updated: Oct 18, 2025

Bioengineering of Humanized Bone Marrow Microenvironments in Mouse and Their Visualization by Live Imaging
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Recreating the Bone Marrow Microenvironment to Model Leukemic Stem Cell Quiescence.

Eimear O'Reilly1, Hojjat Alizadeh Zeinabad1, Caoimhe Nolan1

  • 1Apoptosis Research Centre, Department of Biochemistry, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.

Frontiers in Cell and Developmental Biology
|September 30, 2021
PubMed
Summary

Relapse in acute myeloid leukemia (AML) is driven by quiescent leukemic stem cells (LSCs) protected by the bone marrow microenvironment (BMM). Researchers developed a 3D culture system to model BMM, identifying drug combinations to eradicate these resistant LSCs.

Keywords:
acute myeloid leukemiabone marrow microenvironmentleukemic stem cellquiescencethree-dimensional model

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Area of Science:

  • Hematology
  • Cancer Biology
  • Stem Cell Biology

Background:

  • Relapse remains a significant challenge in acute myeloid leukemia (AML) treatment, primarily due to a persistent population of leukemia-initiating cells (LSCs).
  • Leukemic stem cells (LSCs), similar to long-term hematopoietic stem cells (LT-HSCs), can enter a quiescent state within the bone marrow microenvironment (BMM).
  • This quiescent state confers resistance to conventional therapies by reducing metabolic activity and increasing survival signals, leading to disease re-establishment after treatment cessation.

Purpose of the Study:

  • To develop a reliable experimental model that replicates the quiescence-inducing properties of the bone marrow microenvironment (BMM) for acute myeloid leukemia (AML) cells.
  • To identify therapeutic strategies capable of overcoming BMM-mediated quiescence in leukemic stem cells (LSCs).

Main Methods:

  • A three-dimensional (3D) culture system was established using bone marrow stromal cells, transforming growth factor beta-1, and hypoxia to mimic the BMM.
  • AML cells were cultured in this system, and quiescence was assessed by measuring cell proliferation, cell cycle phase distribution (G0), cell numbers, quiescence marker expression, and metabolic activity.
  • The culture system was adapted into co-axial microbeads to facilitate high-throughput screening.

Main Results:

  • The 3D culture system successfully replicated the quiescence-inducing BMM, leading to a quiescent-like state in AML cells.
  • Confirmed indicators of quiescence included reduced proliferation, increased G0 phase cells, decreased cell numbers, specific marker expression, and diminished metabolic activity.
  • High-throughput screening using the microbead system identified combination drug treatments effective in breaking BMM-mediated LSC quiescence.

Conclusions:

  • A novel 3D culture model effectively recapitulates the bone marrow microenvironment's role in inducing leukemic stem cell (LSC) quiescence.
  • This model provides a platform for identifying therapeutic combinations that can sensitize and eradicate quiescent LSCs, offering a potential strategy to prevent AML relapse.