Abstract
With the aim to detect candidate malignant primitive progenitor populations, we modified an original alkaline phosphatase (ALP) stem cell detection method based on the identification of alkaline phosphatase fluorescent cells in combination with flow cytometry immunophenotyping. Over a period of one year, we have been using this technique to study its activity in patients with leukemia and lymphoma, showing that changes in the alkaline phosphatase levels can be used to detect rare populations of highly refractory malignant cells. By screening different blood cancers, we have observed that this activity is not always restricted to CD34+ leukemic cells, and can be overexpressed in CD34 negative leukemia. We have verified that this method gives accurate and reproducible measurements and our preliminary results suggest that CD34+/ALPhigh cells appear to sustain leukemogenesis over time.
Keywords:
CD34; Vybrant DyeCycle Violet; alkaline phosphatase; leukemic stem cells; stem cells.
MeSH terms
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Adult
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Aged
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Alkaline Phosphatase / metabolism*
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Antigens, CD34 / metabolism
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Antigens, Neoplasm / metabolism
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Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
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Biomarkers, Tumor / metabolism
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Bone Marrow / enzymology
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Bone Marrow / pathology
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Cell Transformation, Neoplastic / metabolism
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Child
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Drug Resistance, Neoplasm
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Early Detection of Cancer / methods
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Enzyme Assays / instrumentation
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Enzyme Assays / methods*
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Female
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Flow Cytometry / instrumentation
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Flow Cytometry / methods
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Gene Rearrangement
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Histone-Lysine N-Methyltransferase / genetics
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Humans
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Immunophenotyping / instrumentation
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Immunophenotyping / methods
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Male
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Myeloid-Lymphoid Leukemia Protein / genetics
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Neoplasm, Residual
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Neoplastic Stem Cells / enzymology*
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Precursor Cell Lymphoblastic Leukemia-Lymphoma / diagnosis*
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Precursor Cell Lymphoblastic Leukemia-Lymphoma / drug therapy
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Precursor Cell Lymphoblastic Leukemia-Lymphoma / enzymology
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Precursor Cell Lymphoblastic Leukemia-Lymphoma / genetics
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Proto-Oncogene Proteins c-myc / genetics
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Reproducibility of Results
Substances
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Antigens, CD34
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Antigens, Neoplasm
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Biomarkers, Tumor
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KMT2A protein, human
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MYC protein, human
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Proto-Oncogene Proteins c-myc
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Myeloid-Lymphoid Leukemia Protein
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Histone-Lysine N-Methyltransferase
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Alkaline Phosphatase