DSMZ no.:

Species:

murine (mouse) (Mus musculus)

Cell type:

hybridoma

Origin:

cells are a subclone derived from B13.29 (first described as clone 9.9) by fusion of BALB/c spleen cells with SP2/0-AG14 myeloma cells; this hybridoma is a non-producer; cells are IL-6-dependent

Reference(s):

Biosafety level:

1

Permissions and restrictions:

Morphology:

round cells growing in suspension; image; image; image

Medium:

80-90% RPMI 1640 + 10-20% h.i. FBS + 50 µM mercaptoethanol + 50-100 pg/ml human IL-6

Subculture:

seed out at ca. 0.3-0.5 x 10⁶ cells/ml, at best in a 24-well plate; split culture 1:2 to 1:6 every 2-3 days; add IL-6 every 1-2 days; cells have a high metabolic activity aciditying the culture medium. CAVEAT: by favoring selective growth, suboptimal culture of cytokine-dependent cell lines may promote outgrowth of factor-independent subclones. Selective conditions include cytokine insufficiency and inadequate cell density. Thus the DSMZ cannot guarantee indefinite stability of factor-dependence of cell lines.

Incubation:

at 37 °C with 5% CO₂

Doubling time:

ca. 20-30 hours

Harvest:

maximal density at about 0.8 x 10⁶ cells/ml

Storage:

frozen with 70% medium, 20% FBS, 10% DMSO

Mycoplasma:

negative in DAPI, microbiological culture, RNA hybridization, PCR assays

Immunology:

bioassay for IL-6 (low cell densities are essential for reliable assays; try doubling cell dilutions beginning with cell densities of 2.5 x 10⁴ cells/ml; duration of assay is commonly 2-3 days)

Species validation:

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.

Cytogenetics:

apparently murine chromosome morphology with extensive centric fusions

Viruses:

ELISA: reverse transcriptase negative; PCR: SMRV -

Supplied as: