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Purification of leukemic blast cells from blood smears using laser microdissection

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Abstract

In treatment of acute myeloid leukemia (AML), prognostic factors, including gene mutation and abnormal gene expression, enable risk stratification of patients. However, in the case of a small proportion of leukemic blast cells, such as AML associated with Down syndrome (AML-DS), it is not possible to examine prognostic factors precisely due to the large proportion of normal cells. Here, we present a novel method for examining prognostic factors by making a smear on a membrane slide glass from a small amount of diagnostic specimen and collecting highly pure leukemic blast cells by laser microdissection (LMD). We verified the effectiveness of this method using 10% KPAM1 cell line suspension and peripheral blood containing 20% blast cells obtained from a patient with transient abnormal myelopoiesis (TAM). After making blood smears, approximately 100 cells were collected and analyzed by direct sequencing. Frameshift mutations (2 bp deletion and 17 bp duplication, respectively) in GATA-1 were detected in each sample, suggesting KPAM1 and TAM blast cells were accurately purified. This novel method enables us to precisely examine prognostic factors in many cases, even in cases with a small proportion of leukemic blast cells or small specimens to preserve.

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Acknowledgements

This work was supported by a Grant-in-Aid for Cancer Research from the Ministry of Health, Labor, and Welfare of Japan (H26-061) and JSPS KAKENHI Grant Number 16K19193.

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Authors and Affiliations

  1. Department of Human Health Sciences, Graduate School of Medicine, Kyoto University, 53 Kawahara-cho, Syogoin, Sakyoku, Kyoto, 606-8507, Japan

    Hidemasa Matsuo, Yasuhiko Kamikubo & Souichi Adachi

  2. Department of Clinical Laboratory, Kyoto University Hospital, Kyoto, Japan

    Hidemasa Matsuo & Shuichi Shiga

  3. Department of Pediatrics, Kurashiki Central Hospital, Kurashiki, Japan

    Tsuyoshi Imai

  4. Department of Pediatrics, Hirosaki University, Hirosaki, Japan

    Tsutomu Toki, Kiminori Terui & Etsuro Ito

Authors
  1. Hidemasa Matsuo
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  2. Shuichi Shiga
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  3. Tsuyoshi Imai
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  4. Yasuhiko Kamikubo
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  5. Tsutomu Toki
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  6. Kiminori Terui
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  7. Etsuro Ito
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  8. Souichi Adachi
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Contributions

Conceived and designed the experiments: HM, TI, YK, TT, KT, EI, and SA. Performed the experiments and analyzed the data: HM, SS, TT, and KT. Wrote the paper: HM.

Corresponding author

Correspondence to Souichi Adachi.

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Conflict of interest

The authors declare that they have no competing interest.

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Matsuo, H., Shiga, S., Imai, T. et al. Purification of leukemic blast cells from blood smears using laser microdissection. Int J Hematol 106, 55–59 (2017). https://doi.org/10.1007/s12185-017-2227-z

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  • DOI: https://doi.org/10.1007/s12185-017-2227-z

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