DSMZ no.:

Species:

human (Homo sapiens)

Cell type:

acute myelomonocytic leukemia

Origin:

established from the peripheral blood of a 29-year-old man with acute myelomonocytic leukemia (AML FAB M4) at diagnosis in 1993; this cell line is TSLP responsive and serves as dendritic cell model. Exome and RNA sequence data are available (see Ref 18187 and Exome sequence and RNA-Seq)

Reference(s):

14618, 14482, 14919, 14918, 14722, 14739, 18187, 16556, 16670, 40552

Biosafety level:

1

Permissions and restrictions:

Depositor-specific restrictions:

Transfer to third parties needs to be negotiated with the depositor.

Morphology:

round blast-like cells growing as single cells and sometimes in small clumps in suspension; round cells are in active growth phase and should be in the majority; image; image

Medium:

60% alpha-MEM (with ribo- and deoxyribonucleosides) + 20% h.i. FBS + 20% vol conditioned medium of cell line 5637 (DSMZ ACC 35); according to ref 14618 proliferation of this cell line was described also in the presence of GM-CSF (100 U/ml) and IL-3 (100 U/ml) and M-CSF (100 U/ml) or SCF (50 ng/ml)

Subculture:

cultivate in 24-well-plates (strongly recommended); seed out at ca. 1.0 x 10⁶ cells/ml/well of a 24-well plate; viability may decrease within the first days and growth may start only 1-2 weeks after thawing; in growth phase split 1:2 to 1:3 every 4-6 days; at feeding or splitting about 1/4 of the spent medium should be retained; maintain at 0.4-1.0 x 10⁶ cells/ml

Incubation:

at 37 °C with 5% CO₂

Doubling time:

about 100-120 hours after thawing; in optimal conditions 50-70 hours

Harvest:

saturation density at 1.5-2.0 x 10⁶ cells/ml

Storage:

frozen with 70% medium, 20% FBS, 10% DMSO

Mycoplasma:

negative in DAPI, microbiological culture, RNA hybridization, PCR assays

Immunology:

CD3-, CD4(+), CD5-, CD7-, CD8-, CD13, CD14-, CD15(+), CD16-, CD19-, CD33+, CD34(+), HLA-DR(+), cyCD3-, cyCD68(+); image

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 near-diploid karyotype with 6% tetraploidy - 46(44-48)<2n>XY, t(1;3)(q43;q13)inv(3)(q21q26), t(2;7)(q36;q36)inv(7)(p15q36), t(12;22)(p13;q12) - carries t(12;22) recurrent in AML M4 - also carries masked inv(3) associated with abnormal megakaryocytopoiesis

Viruses:

ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HHV-8 -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV -

Supplied as: