0175 - M-NFS-60

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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: It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
  1. Thaw the vial by gently agitating it in a 37°C water bath. To minimize contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as its contents are thawed and decontaminate it by dipping in or spraying with 70% ethanol. From this point, all operations must be performed under strict aseptic conditions.
  3. For cells sensitive to DMSO, it is recommended to remove the cryoprotective agent immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL of complete culture medium and centrifuge at approximately 125 × g for 5 to 7 minutes.
  4. Discard the supernatant and resuspend the cell pellet in the recommended complete medium (see specific batch information for the appropriate dilution ratio).
  5. Incubate the culture under appropriate atmospheric and temperature conditions (see "Culture Conditions" for this cell line).

NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at 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
Cellosaurus