DSMZ no.:

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)

Reference(s):

14980, 40446, 40447, 18187

Biosafety level:

1

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 in serum-free freezing medium (Bambanker^TM)

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(+), CD80+, CD138(+), HLA-DR+, cyCD79a+, cIgG-, cIgM+, sIg-kappa-, sIg-lambda-; 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: