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BCRJ Code 0175
Cell Line M-NFS-60
Species Mus musculus
Vulgar Name Mouse
Cell Type Virus Induced
Morphology Lymphoblast
Disease Myelogenous Leukemia
Growth Properties Suspension
Derivation The M-NFS-60 cell line was derived from a myelogenous leukemia induced with the Cas-Br-MuLV wild mouse ecotropic retrovirus.
Applications This cell line is a suitable transfection host.
Biosafety 1
Addtional Info The cells are responsive to both interleukin 3 (interleukin-3, IL-3) and macrophage colony stimulating factor (M-CSF). The cells contain a truncated c-myb proto - oncogene caused by integration of a retrovirus.
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose, 0.05 mM 2-mercaptoethanol, 62 ng/ml human recombinant macrophage colony stimulating factor (M-CSF) and 10% of fetal bovine serum.
Subculturing Cultures can be maintained by addition or replacement of fresh medium. Subculture every two days at 2.5 X 10e4 viable cells/mL.
Subculturing Medium Renewal Add fresh medium at the time of subculture
Culture Conditions Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
References 22842: Nakoinz I, et al. Differentiation of the IL-3-dependent NFS-60 cell line and adaption to growth in macrophage colony-stimulating factor. J. Immunol. 145: 860-864, 1990. PubMed: 2142710 23295: Weinstein Y, et al. Truncation of the c-myb gene by a re
Depositors Cristina Massoco; Ciallyx