Species:
human (Homo sapiens)
Cell type:
chronic B cell leukemia
Origin:
established in 1993 from the peripheral blood of a 61-year-old man with chronic B cell leukemia (B-CLL in prolymphocytoid transformation to B-PLL); serial sister cell line of MEC-2 (DSMZ ACC 500). 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:
Morphology:
round to polymorphic cells growing in suspension, singly or partly in small aggregates, a few cells are slightly adherent;
image;
image
Medium:
90% Iscove's MDM + 10% h.i. FBS
Subculture:
seed out at ca. 0.5 x 106 cells/ml; after thawing, during the first week cells should be kept with 20% FBS initially; culture in 24-well-plate is of advantage; maintain at about 0.5-2.0 x 106 cells/ml; split saturated culture 1:2 to 1:3 every 2-3 days
Incubation:
at 37 °C with 5% CO2
Doubling time:
ca. 40 hours
Harvest:
cell harvest of about 2.0 x 106 cells/ml
Storage:
frozen with 70% medium, 20% FBS, 10% DMSO
Mycoplasma:
initial contamination was eliminated with BM-Cyclin (tiamulin & minocycline), then negative in microbiological culture, RNA hybridization, and continuously in PCR assays
Immunology:
CD3 -, CD10 -, CD13 -, CD19 +, CD20 +, CD34 -, CD37 +, CD38 +, cyCD79a +, CD80 +, CD138 +, HLA-DR +;
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, identical to sister cell line MEC-2 (ACC 500)
Cytogenetics:
human near-diploid karyotype with 10% polyploidy - 46(44-47)<2n>XY, -2, +7, -12, +1-2mar, t(1;6)(q22-23;p21), add(7)(q11), der(10)(10pter->q22::?::2q11->qter), del(17)(p11) - small acf/mar present in most cells - resembles published karyotype
Viruses:
PCR: EBV +, HBV -, HCV -, 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
