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Updated: Mar 15, 2026
Flow Cytometry to Estimate Leukemia Stem Cells in Primary Acute Myeloid Leukemia and in Patient-derived-xenografts, at Diagnosis and Follow Up
Published on: March 26, 2018
Immunological Analyses of Leukemia Stem Cells
Kazuhito Naka1, Yoshihiro Takihara2
1Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8553, Japan. kanaka55@hiroshima-u.ac.jp.
Insights
New Duolink(®) in situ PLA technology offers a quantitative method to study proteins in rare CML Leukemia stem cells (LSCs). This approach enhances understanding of LSC biology and may lead to new CML therapies.
Area of Science:
- Hematology
- Molecular Biology
- Cell Biology
Background:
- Assessing protein characteristics in Chronic Myeloid Leukemia (CML) Leukemia stem cells (LSCs) traditionally relied on fluorescence immunohistochemistry (FIHC).
- FIHC has limitations in quantifying specific molecular events within rare cell populations.
Purpose of the Study:
- To introduce and evaluate Duolink(®) in situ Proximity Ligation Assay (PLA) as a novel, quantitative method for studying CML LSCs.
- To explore the potential of in situ PLA for analyzing signal transduction, posttranslational modifications, and protein-protein interactions at the single-cell level in CML LSCs.
Main Methods:
- Utilized Duolink(®) in situ PLA technology, which employs two antibody-based probes.
- Applied the method to analyze protein characteristics in rare CML LSC populations.
Main Results:
- Duolink(®) in situ PLA provides a more quantitative approach compared to traditional methods.
- The technology enables detailed analysis of molecular events at the single-stem-cell level.
Conclusions:
- Duolink(®) in situ PLA significantly advances the understanding of CML LSC biology.
- This methodology holds promise for the development of targeted therapeutics to eradicate CML LSCs.
Abstract:
Traditionally, the intracellular localization and expression levels of specific proteins in CML Leukemia stem cells (LSCs) have been evaluated by fluorescence immunohistochemistry (FIHC). More recently, Duolink(®) in situ PLA technology has opened up a new and more quantitative way to evaluate signal transduction, posttranslational modification, and protein-protein interaction at the single-stem-cell level. This novel methodology, which employs two antibody-based probes, has already increased our understanding of the biology of the rare CML LSC population. In the future, the use of this approach may contribute to the development of novel therapeutics aimed at eradicating CML LSCs in CML patients.
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