Species:
human (Homo sapiens)
Cell type:
erythroleukemia
Origin:
established from the bone marrow of a 35-year-old man with erythroleukemia (AML M6) in 1987; cell line was first called MFD-1; cells are described in the literature to be proliferatively responsive to several hematopoietic growth factors including EPO, GM-CSF, IFN-gamma, IL-3, IL-4, IL-5, IL-6, IL-13, LIF, NGF, OSM, SCF, and TNF-alpha (see Mire-Sluis et al., J. Immunol. Methods 187: 191-199, 1995); we have noted that the cell line can become very quickly cytokine-independent (within 3-4 weeks), presumably due to outgrowth of independent cells. Exome and RNA sequence data are available (see Ref 18187 and
Exome sequence and
RNA-Seq)
Permissions and restrictions:
Morphology:
large, single, round cells in suspension; cells shed large amounts of cytoplasmatic particles that accumulate in the culture (not to be confused with bacteria);
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Medium:
60-70% RPMI 1640 + 20% h.i. FBS + 10-20% vol conditioned medium of cell line 5637 (DSM ACC 35) (or 1-5 ng/ml recombinant GM-CSF or IL-3)
Subculture:
seed out at ca. 1 x 106 cells/ml in a 24-well-plate or a small culture flask; split saturated culture 1:2 every 3 days; optimal density at ca. 0.5-1.0 x 106 cells/ml. CAVEAT: by favoring selective growth, suboptimal culture of cytokine-dependent cell lines may promote outgrowth of factor-independent subclones. Selective conditions include cytokine insufficiency and inadequate cell density. Thus the DSMZ cannot guarantee indefinite stability of factor-dependence of cell lines.
Incubation:
at 37 °C with 5% CO2
Doubling time:
ca. 70 hours
Harvest:
maximal density at ca. 1.3-1.5 x 106 cells/ml
Storage:
frozen with 70% medium, 20% FBS, 10% DMSO
Mycoplasma:
initial contamination was eliminated with BM-Cyclin (tiamulin & minocycline), then negative in DAPI, microbiological culture, RNA hybridization, and continuously in PCR assays
Immunology:
CD3 -, CD13 +, CD14 -, CD15 -, CD19 -, CD33 +, CD34 +, CD71 +, HLA-DR +;
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Authenticity:
STR analysis according to the global standard ANSI/ATCC ASN-0002.1-2021 (2021) resulted in an authentic STR profile of the reference STR database.
Cytogenetics:
human highly rearranged hyperdiploid karyotype with 12% polyploidy - 52-57<2n>XY/XXY, +3, +5, +6, -8, +12, +15, +19, +19, +20, +20, +3mar, der(1)?dup(1)(p21p31)t(1;8)(p36;q11), t(2;12)(q32;q14), t(3;12)(p13-14;p12-13), add(3)(q21), add(5)(q11-13), der(8)t(1;8)(p36;q11), der(12)t(3;12)(p13-14;p12-13)t(1;12)q31-32;q24), add(14)(p12), iso(17)(q10)add(17)(q21), add(19)(q13), trp(19)(q12;q13.3), der(21)t(19;21)(q13.1;q22)dup(19)(q13.1q13.3)t(11;19)(q13;q13.3), der(22)t(19;22)(q11;p11) - resembles published karyotype
Viruses:
ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HHV-8 -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV -
Supplied as:
| Delivery form | | Prices |
|
| Frozen culture | | 450,- € |
Growing culture
(please inquire for exact delivery time) | | 900,- € |
| DNA isolated from cell line (25 µg) | | 560,- € |
| DNA isolated from cell line (5 µg) | | 140,- € |
see price list
