NFS-60 cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

NFS-60 cell line细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
Designation: NFS-60

Origin and General Characteristics
Organism: Mus musculus (mouse)
Tissue: Blood
Morphology: Lymphoblast
Cell type: Leukemia, myeloid
Growth Properties: Suspension

Description: A murine myeloblastic cell line established from leukemic cells obtained after infection of
(NFS X DBA/2) F1 adult mice with Cas Br-M murine leukemia virus. NFS-60 cells are
dependent on IL3 for growth and maintenance of viability in vitro. These cells are used to
assay murine and human G-CSF. This bipotential murine hematopoietic cell line is
responsive to IL-3, GM-CSF, G-CSF, and erythropoietin.

Culture Conditions and Handling
Culture Medium: RPMI 1640 medium supplemented with L-glutamine, 1 mM Na-pyruvate, 10% fetal
bovine serum and IL-3. The appropriate concentration of the IL-3 depends on the source
and quality and must be determined by the user. As source of cytokines, conditioned
medium supplement (KMG-2, order number 810210, 25ml, 810250,
50ml), 10-20% in regular culture medium may be used as an alternative. The Ready-touse
medium, MG-71, incl. KMG-2, is available by the order number 820701.
Subculturing: Subculture by transferring an appropriate amount of the cell suspension into new cell
culture flasks already containing fresh cell culture media. Start cultures at 5 x 104 viable
cells/ml.

Freeze Medium: CM-2, a chemically defined, serumfree
cryomedium.

Sterility: Fluorescence (DAPI) test: negative; Mycoplasma specific PCR: negative; Bacteria
specific PCR: negative

Biosafety Level: 1

Special Features of the Cell Line
Species identification: Mouse origin was verified by the PCR technique using the Mouse cox I and Mouse
J01420 primer.

Viruses: SMRV: Negative, as confirmed by Real-Time PCR

References:
Weinstein Y et al. Truncation of the c-myb gene by a retroviral integration in an interleukin 3-dependent myeloid
leukemia cell line. Proc Natl Acad Sci USA 83: 5010-4, 1986.

Recommendations for handling of suspension cells following delivery
Cryopreserved cells
If immediate culturing is not intended, the cryovial(s) may be stored in liquid nitrogen after arrival.
If immediate culturing is intended, please follow these instructions:
Quickly thaw by rapid agitation in a 37°C water bath within 40-60 seconds. The water bath should have clean
water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water
bath (a small ice clump should remain and the cryovial should still be cold).
From now on, all operations should be carried out under aseptic conditions.
Immediately transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and
transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium. Resuspend the cells
carefully. The cells should be spun down at 300xg for 3 minutes. After centrifuging, aseptically remove the
supernatant and add 10 ml of fresh cell culture media. Carefully resuspend the cells and distribute into one 25cm2
cell culture flask. Incubate at 37°C/5% CO2.
Subculture as soon as the cell concentration has reached 1 x 106 cells/ml. It is recommended to distribute the
cells into new flasks containing fresh medium thus diminishing the amount of dead cells and cell debris. Adjust to
a cell concentration of 1-2 x 105 cells/ml depending on the specification given for the cell line.
After about 1-2 times of sub-culturing as recommended the percentage of viable cells should be > 90%.
Proliferating Cultures
Immediately after receipt the cell concentration should be determined. If the cell concentration already has
reached a value of 1 x 106 cells/ml or even more, subculture the cells as described above. Remove the entire
content of the flask and centrifuge at 300xg for 10 minutes.
Resuspend the cell pellets as suggested under subculture procedures described on the appropriate datasheet.

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