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| 产品名称： | CRO-AP2 |
|---|---|
| 商品货号： | TS258850 |
| <strong>Cell line </strong>： | CRO-AP2 |
| DSMZ no. ： | TS258850 |
| Species ： | confirmed as human with IEF of AST, LDH, MDH |
| Cell type ： | B cell lymphoma |
| Origin ： | established in 1996 from the diagnostic pre-treatment pleural effusion sampled preterminally of a 49-year-old HIV+ man with primary effusion lymphoma (PEL) and previous history of Kaposis sarcoma; autopsy revealed involvement of serous membranes without 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) |
| Reference(s) ： | 15032, 14417, 14483, 18187 |
| Biosafety level ： | 2 |
| 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 ： | A, B |
| <strong> </strong>： | DSMZ Scientific Data: |
| 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 |
| Fingerprint ： | multiplex PCR of minisatellite markers revealed a unique DNA profile |
| 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-I/II -, MLV -, SMRV - |