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Modeling Chemotherapy Resistant Leukemia In Vitro
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Modeling Chemotherapy Resistant Leukemia In Vitro.

William L Slone1, Blake S Moses1, Rebecca Evans1

  • 1Alexander B. Osborn Hematopoietic Malignancy and Transplantation Program of the Mary Babb Randolph Cancer Center, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine.

Journal of Visualized Experiments : Jove
|February 19, 2016
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Summary

Researchers developed a new method to isolate therapy-resistant leukemic cells from the bone marrow microenvironment. This technique helps in studying chemoresistance and developing new treatments for minimal residual disease.

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Area of Science:

  • Hematology
  • Cancer Biology
  • Cell Biology

Background:

  • The bone marrow microenvironment is a sanctuary for hematopoietic diseases.
  • Leukemic cells can initiate and progress within the bone marrow niche.
  • Understanding interactions between leukemic cells and the bone marrow niche is crucial for effective treatment.

Purpose of the Study:

  • To develop a reliable in vitro model for studying chemoresistance in leukemic cells.
  • To isolate therapy-resistant leukemic cells from co-cultures with bone marrow stromal cells and osteoblasts.
  • To provide a source of purified resistant cells for investigating resistance mechanisms and testing novel therapies.

Main Methods:

  • Co-culture of human leukemic cells with primary human bone marrow stromal cells (BMSC) and osteoblasts (OB).
  • Strict feeding schedule to control nutrient availability and prevent nutrient fluxes.
  • Gel type 10 cross-linked dextran (G10) particle-based recovery of leukemic cells that migrated beneath adherent BMSC or OB, forming a 'phase dim' (PD) population.

Main Results:

  • The co-culture system generated a chemoresistant subpopulation of leukemic cells.
  • The G10 particle method successfully isolated a purified 'phase dim' (PD) population of therapy-resistant leukemic cells.
  • This PD population represents a clinically relevant model for studying resistance mechanisms.

Conclusions:

  • The described in vitro model provides a consistent source of purified, therapy-resistant leukemic cells.
  • This model enables rigorous investigation of apoptotic, metabolic, and cell cycle pathways involved in chemoresistance.
  • The isolated PD cells serve as a valuable tool for pre-clinical testing of novel therapeutic strategies targeting minimal residual disease.