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December 8, 2025BloodOpen Access

A defined culture model for ex vivo expansion of primary AML cells preserves leukemic stem cells and clonal architecture for functional and therapeutic studies

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Authors

UGUpendar R. GollaKMKentaro MinagawaHZHong Zheng

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Overview

Ex vivo culture of primary AML cells maintains leukemic stem cells and CD11b differentiation, suggesting therapeutic avenues with Venetoclax.

Key Points

  • To present a defined culture model for the ex vivo expansion of primary AML cells while preserving leukemic stem cells and clonal architecture.
  • Utilized a serum-free culture system with cytokines for primary human AML cells from blood/bone marrow (n=30)
  • Cultured cells under five conditions with varying cytokines and small molecules
  • Assessed stemness and differentiation via flow cytometry
  • Evaluated transplantability of cultured cells in NRG-S mice
  • Performed NGS on AML cases to compare mutational profiles after culture.
  • Condition C5 showed the highest expansion of 389-fold; C2 maintained best CD34% stem cell levels
  • C2 and C3 conditions selected for further studies due to better engraftment rates in mice
  • NGS revealed stable mutational allele frequencies in cultured AML cells
  • Venetoclax induced apoptosis and affected CD34+ stemness.

Cite This Study

Golla et al. (2025) studied this question.

synapsesocial.com/papers/69362f574fa91c937236d9fehttps://doi.org/10.1182/blood-2025-3488
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