MEC-1 cell line慢性B细胞白血病细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

MEC-1 cell line慢性B细胞白血病细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
Cell line: MEC-1 DSMZ no.: ACC 497Species: human (Homo sapiens)Cell type: chronic B cell leukemia Origin: established in 1993 from the peripheral blood of a 61-year-old Caucasian man with chronic B cell leukemia (B-CLL in prolymphocytoid transformation to B-PLL); serial sister cell line of MEC-2 (DSMZ ACC 500) Reference(s): 14980Biosafety 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 Commission for Biosafety (ZKBS) the cells have to be handled under biosafety level 2 when the cells are used for genetic manipulations. Permissions and restrictions: A  DSMZ Cell Culture Data: Morphology:round to polymorphic cells growing in suspension, singly or partly in small aggregates, a few cells are slightly adherent; imageMedium: 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  DSMZ Scientific Data: Mycoplasma: contamination was eliminated with BM-Cyclin (tiamulin & minocycline), then negative in microbiological culture, RNA hybridization, PCR assays Immunology: CD3 -, CD10 -, CD13 -, CD19 +, CD20 +, CD34 -, CD37 +, CD38 +, cyCD79a +, CD80 +, CD138 +, HLA-DR +; image Fingerprint: same DNA profile as MEC-2 using multiplex PCR of minisatellite markers Species: confirmed originally as human by cytogenetics 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-I/II -, MLV -, SMRV
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