Subculture:
seed out at ca. 0.4-0.5 x 106 cells/ml; split saturated culture 1:2 every 3 days; cells may grow slightly adherent; cells may be detached by tapping the flask or with a rubber policeman or with TNE (40 mM TrisCl pH 7.5, 1 mM EDTA pH 7.5, 150 mM NaCl); trypsin/EDTA usually not necessary
Mycoplasma:
negative in DAPI, microbiological culture, RNA hybridization, PCR assays
Immunology:
CD3 -, CD5 -, CD10 -, CD13 -, CD19 -, CD20 -, CD34 -, CD38 +, CD80 -, CD138 +, sm/cyIgG -, sm/cyIgM -, sm/cykappa -, sm/cylambda -;
image
Cytogenetics:
human flat-moded hypotriploid karyotype with 7.5% polyploidy - 62-67<3n>XXY, -1, +5, +6, +7, -8, -9, -10, +11, -12, -13, -14, +16, +18, -19, +21, -22, der(1)t(1;1)(p36.3;q11-12), add(3)(q27-29)x2, del(4)(p14p15.4), der(5;6)(q10;p10)x1-2, del(5)(q32), del(6)(q15), add(9)(p24), add(11)(p15), del(11)(q25), der(14)t(1;14)(q11-12;q32)x2, del(15)(q11.2q14)x2, add(16)(q23-24), der(17)t(?8;17)(q21.2;q25), del(22)(q13.2) - der(1), 6q-, 14q+ and 22q- associated with multiple myeloma - identified by disomic 14q+
Viruses:
ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV -
