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This is a leukemic cell line with an 8;21 chromosome translocation. This translocation juxtaposes the AML1 with ETO (or MTG8) gene, giving rise to the fusion gene AML1-ETO (also known as AML1-MTG or RUNX1-CBF2T1), hence the cells produce chimeric AML1-ETO protein. This protein down-regulates CEBPA mRNA, protein and DNA binding activity, which is crucial for the differentiation of granulocytes. The cell line was established from the peripheral blood of an acute myeloid leukemia (AML) patient. The cells are positive for myeloperoxidase showing a morphology of myeloid maturation. In proliferation assay the cells in culture showed response to interleukin-3 (IL-3), IL-6, granulocyte colony-stimulating factor (G-CSF), and granulocytemacrophage CSF (GM-CSF), but not to IL-1 or IL-5. Neither granulocytic nor eosinophilic maturation was observed in the in vitro liquid culture by the addition of dimethyl sulfoxide, G-CSF, or IL-5, respectively. Induction of macrophagelike cells was seen by the addition of phorbol ester. Proliferation is inhibited by 1,25S-(OH)2-16,23-diene-26-F3-10-nor D3.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 20%. Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Subculturing:
Protocol: Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 3 X 10(5) viable cells/ml. Interval: Maintain cell density between 3 X 10(5) and 3 X 10(6) viable cells/ml. Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Doubling Time:
40 to 48 hrs
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2001
recommended serum:ATCC 30-2020
57589: Tashiro S, et al. Establishment of a human acute myeloid leukemia cell line (Kasumi-1) with 8;21 chromosome translocation. Blood 77: 2031-2036, 1991. PubMed: 2018839 61289: Miyoshi H, et al. The t(8;21) translocation in acute myeloid leukemia results in production of an AML1-MTG8 fusion transcript. EMBO J. 12: 2715-2721, 1993. PubMed: 8334990 61291: Miyoshi H, et al. t(8;21) breakpoints on chromosome 21 in acute myeloid leukemia are clustered within a limited region of a single gene, AML1. Proc. Natl. Acad. Sci. USA 88: 10431-10434, 1991. PubMed: 1720541 61292: Asou H, et al. 19-nor vitamin-D analogs: a new class of potent inhibitors of proliferation and inducers of differentiation of human myeloid leukemia cell lines. Blood 92: 2441-2449, 1998. PubMed: 9746784 61293: Kozu T, et al. Junctions of the AML1/MTG8(ETO) fusion are constant in t(8;21) acute myeloid leukemia detected by reverse transcription polymerase chain reaction. Blood 82: 1270-1276, 1993. PubMed: 8353289 61295: Pabst T, et al. AML1-ETO downregulates the granulocytic differentiation factor C/EBPalpha in t(8;21) myeloid leukemia. Nat. Med. 7: 444-451, 2001. PubMed: 11283671
