Abstract
Using serum-containing culture, we examined whether AGM-S3 stromal cells, alone or in combination with hematopoietic growth factor(s), stimulated the proliferation of CD34(+) cells from patients with juvenile myelomonocytic leukemia (JMML). AGM-S3 cells in concert with stem cell factor plus thrombopoietin increased the numbers of peripheral blood CD34(+) cells to approximately 20-fold of the input value after 2 weeks in nine JMML patients with either PTPN11 mutations or RAS mutations, who received allogeneic hematopoietic transplantation. Granulocyte-macrophage colony-stimulating factor (GM-CSF) also augmented the proliferation of JMML CD34(+) cells on AGM-S3 cells. The expansion potential of CD34(+) cells was markedly low in four patients who achieved spontaneous hematological improvement. A large proportion of day-14-cultured CD34(+) cells were negative for CD38 and cryopreservable. Cultured JMML CD34(+)CD38(-) cells expressed CD117, CD116, c-mpl, CD123, CD90, but not CXCR4, and formed GM and erythroid colonies. Day-7-cultured CD34(+) cells from two of three JMML patients injected intrafemorally into immunodeficient mice stimulated with human GM-CSF after transplantation displayed significant hematopoietic reconstitution. The abilities of OP9 cells and MS-5 cells were one-third and one-tenth, respectively, of the value obtained with AGM-S3 cells. Our culture system may provide a useful tool for elucidating leukemogenesis and for therapeutic approaches in JMML.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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ADP-ribosyl Cyclase 1 / genetics
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ADP-ribosyl Cyclase 1 / metabolism
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Adolescent
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Animals
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Antigens, CD34 / genetics
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Antigens, CD34 / metabolism
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Cell Proliferation / drug effects
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Clone Cells
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Coculture Techniques
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Embryonic Stem Cells / drug effects*
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Embryonic Stem Cells / metabolism
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Embryonic Stem Cells / pathology
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GTP Phosphohydrolases / genetics
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GTP Phosphohydrolases / metabolism
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Gene Expression Regulation, Leukemic*
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Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology*
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Hematopoietic Stem Cells / drug effects*
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Hematopoietic Stem Cells / metabolism
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Hematopoietic Stem Cells / pathology
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Humans
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Leukemia, Myelomonocytic, Juvenile / genetics*
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Leukemia, Myelomonocytic, Juvenile / metabolism
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Leukemia, Myelomonocytic, Juvenile / pathology
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Mice
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Mice, Inbred NOD
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Mice, SCID
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Mutation
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Neoplastic Stem Cells / metabolism
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Neoplastic Stem Cells / pathology
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Neoplastic Stem Cells / transplantation
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Protein Tyrosine Phosphatase, Non-Receptor Type 11 / genetics
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Protein Tyrosine Phosphatase, Non-Receptor Type 11 / metabolism
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins p21(ras)
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Signal Transduction
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Stromal Cells / drug effects*
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Stromal Cells / metabolism
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Stromal Cells / pathology
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ras Proteins / genetics
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ras Proteins / metabolism
Substances
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Antigens, CD34
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KRAS protein, human
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Membrane Proteins
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Proto-Oncogene Proteins
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Granulocyte-Macrophage Colony-Stimulating Factor
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PTPN11 protein, human
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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ADP-ribosyl Cyclase 1
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GTP Phosphohydrolases
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NRAS protein, human
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Proto-Oncogene Proteins p21(ras)
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ras Proteins