Species:
human (Homo sapiens)
Cell type:
chronic B cell leukemia
Origin:
established from the peripheral blood of a 73-year-old man with B-prolymphocytic leukemia (PLL) at diagnosis; cell line was established by EBV-transformation during treatment with phorbol ester TPA; cells express mRNA of the proto-oncogenes BCL2 and BCL3. 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 and thus the cell line is categorized biosafety level 1. According to a recommendation of the German Central Committee for Biological Safety (ZKBS) the cells have to be handled under biosafety level 2 when the cells are used in genetic engineering projects.
Permissions and restrictions:
Medium:
90% RPMI 1640 + 10% h.i. FBS
Subculture:
seed out at ca. 0.5 x 106 cells/ml; maintain at 0.1-0.5 x 106 cells/ml; split saturated culture 1:3 to 1:4 every 3-4 days
Incubation:
at 37 °C with 5% CO2
Doubling time:
ca. 30-40 hours
Harvest:
maximum density at 1.0-1.5 x 106 cells/ml
Storage:
frozen with 70% medium, 20% FBS, 10% DMSO
Mycoplasma:
initial contamination was eliminated with Ciprobay (ciprofloxacin), then negative in DAPI, microbiological culture, RNA hybridization, and continuously in PCR assays
Immunology:
CD3 -, CD10 -, CD13 -, CD19 +, CD20 +, CD34 -, CD37 +, cyCD79a +, CD80 +, 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 near tetraploid karyotype with clear mode - 87-93<4n>XXYY, +3, -7, -8, -10, +12, +12, -14, -19, -19, -20, -20 - appears to be derivate of original hyperdiploid line which exhibited trisomy 12
Molec. Genetics:
exome sequencing revealed a BRAF K601N mutation
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
