Species:
human (Homo sapiens)
Cell type:
B cell lymphoma
Origin:
established in 1996 from the diagnostic pre-treatment pleural effusion sampled preterminally of a 49-year-old man with primary effusion lymphoma (PEL) and previous history of Kaposi's sarcoma; autopsy revealed involvement of serous membranes with formation of solid tumor mass; cells were described in the literature to be EBV+, HHV-8+ and to carry a BCL6 point mutation. 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. However, expression of immediate-early protein BZLF-1 and lately expressed capsid protein were negative by western blot and immunostaining, respectively, in untreated and phorbol ester / sodium butyrate stimulated cells. The infection was classified as latent infection without production of active viruses. A transmission of EBV during handling of the cells is unlikely.
Additionally, the cell line carries HHV-8 sequences which were detected by PCR and Southern blot. We did not determine the infection mode of this herpes virus and cannot exclude the secretion of the viruses. The cell line is thus categorized biosafety level 2.
Permissions and restrictions:
Morphology:
lymphoblast-like cells growing in clumps or as single cells in suspension;
image;
image
Medium:
80% RPMI 1640 + 20% h.i. FBS
Subculture:
seed out after thawing at ca. 0.5 x 106 cells/ml; split saturated culture about 1:2 to 1:3 every 2-4 days; maintain at ca. 0.2-0.5 x 106 cells/ml feeding cultures daily; viability decreases dramatically after thawing, however cells recover within 1-2 weeks but will not exceed 80%; cells are sensitive to freezing methods, best results are obtained with a constant cooling rate to -196°C; cooling to -80°C applying a box with isopropanol and subsequent transfer to liquid nitrogen is not recommended
Incubation:
at 37 °C with 5% CO2
Doubling time:
ca. 3-4 days
Harvest:
maximal density at ca. 1.5 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 -, CD10 -, CD13 -, CD19 -, CD20 -, CD34 -, CD38 +, CD80 -, CD138 +, HLA-DR +, sm/cyIgG -, sm/cyIgM -, sm/cykappa -, sm/cylambda -;
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 hyperdiploid karyotype with 12% polyploidy - 48(47-48)<2n>X, +X, -Y, +12, add(?Y)(p11), add(1)(q43.1), der(15)t(8;15)(q11;q25), der(22)t(1;22)(q21;q13) - sideline with add(9)(q13) - 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
