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described as secreting a mouse monoclonal antibody (IgG2a) CD2; the cell line was established in 1987 by fusion of PX63AG8.653 cells with spleen cells of a mouse immunized with fresh peripheral blood lymphocytes from patients with expansions of gamma-delta Tcells
80% Dulbecco's MEM (4.5 g/L glucose) + 20% h.i. FBS + HFCS ("Hybridoma Fusion and Cloning Supplement")
split saturated cultures 1:2 to 1:3 every 3 days; seed out at 0.8 x 106 cell/ml, best in 12-well-plate with HFCS; maintain at 0.3-0.5 x 106 cells/ml; some cells attach to the surface of the culture flask, but can be detached by tapping the flask; for the first 2 weeks, HFCS should be used which thereafter can be reduced to 50% or 25% of the recommended concentration; cells grow very slowly in the complete absence of HFCS
at 37°C with 10% CO2
about 40-50 hours
maximal density of about 1 x 106 cells/ml
frozen with 70% medium, 20% FBS, 10% DMSO
DSMZ Scientific Data:
negative in DAPI, microbiological culture, RNA hybridization, PCR assays
secretion of a IgG2a monoclonal antibody
Species-level identification was carried out by mitochondrial Cytochrome C Oxidase Subunit 1 (COI) DNA barcoding according standard ANSI/ATCC ASN-0003-2015 and revealed Mus musculus species.