Species:
human (Homo sapiens)
Cell type:
T/NK cell leukemia
Origin:
established from the pericardial fluid of a 15-year-old man with acute lymphoblastic leukemia (ALL) at relapse (with accompanying thymoma) in 1983; cells were described as: (i) showing NK activity and ADCC (antigen-dependent cell-mediated cytotoxicity), (ii) having their T cell receptor (TCR) genes in germline configuration, and (iii) expressing no TCR proteins (also CD3-, CD16-, CD56+, CD57-). Exome and RNA sequence data are available (see Ref 18187 and
Exome sequence and
RNA-Seq)
Risk assessment:
The cell line is positive for EBV (HHV-4) by PCR analysis. Additionally, expression of immediate-early protein BZLF-1 and lately expressed capsid protein were also positive by western blot and immunostaining, respectively, in untreated and phorbol ester / sodium butyrate stimulated cells. The infection was classified as lytic associated with a production of active viruses. A transmission of EBV during handling of the cells is possible and the cell line is thus categorized biosafety level 2.
Permissions and restrictions:
Morphology:
round to polygonal cells growing singly or in clumps in suspension;
image;
image;
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Medium:
80% Iscove's MDM + 20% h.i. FBS
Subculture:
start culture in a 24-well-plate and seed out at ca. 0.8-1.5 x 106 cells/ml, initially after thawing the viability may be reduced down to ca. 20-50% during the first 1-2 weeks (during that initial period, cells should not be diluted, medium may be exchanged by centrifugation); in their vigorous growth phase, maintain at 0.2-0.5 x 106 cells/ml and split saturated culture 1:2 every second day; a significant amount of cell debris may allways be present in the culture
Incubation:
at 37 °C with 5% CO2
Doubling time:
ca. 40-50 hours
Harvest:
cell harvest of ca. 0.6 x 106 cells/ml
Storage:
frozen with 70% medium, 20% FBS, 10% DMSO
Mycoplasma:
negative in DAPI, microbiological culture, RNA hybridization, PCR assays
Immunology:
CD2 -, CD3 -, CD4 -, CD5 -, CD6 -, CD7 +, CD8 -, CD13 -, CD19 -, CD25 (+), CD30 +, CD34 -, TCRalpha/beta -, TCRgamma/delta -;
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 flat-moded near-tetraploid karyotype with 12% polyploidy - 84-98<4n>XXYY, -6, -9, -15, -19, +6-8mar, der(X)t(X;7)(q25;q21), der(1)dup(1)(q12.2q2?2)t(1;17)(q3?1;q1?2), der(4)t(1;4)(q32.2;q35)x2, der(5)t(X;5)(p1?3;q21)x2, del(6)(p11), der(10;17)(q10;q10), dup(14)(q1?q2?), add(16)(q24), add(17)(q11) - resembles published karyotype
Viruses:
ELISA: reverse transcriptase negative; PCR: EBV +, HBV -, HCV -, HHV-8 -, HIV -, 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
