Product category

Human cells

Synonyms

SK_N_FI; SKNFI; SK-N-F1; SKNF1

Organism

Homo sapiens, human

Cell type

neuroblast

Morphology

epithelial

Tissue

Brain

Disease

Neuroblastoma

Applications

3D cell culture

Immunology

Neuroscience

Growth properties

Adherent

Derivation

SK-N-FI is a neuroblastoma cell line derived in 1979 from a bone marrow metastasis from a 11 year old Caucasian male with poorly differentiated embryonal neuroblastoma.

Population doubling time

-

Age

11 years

Ethnicity

White

Gender

Male

Tumorigenic

Yes;

Yes, forms tumors in nude mice with a latent period of 4 to 6 weeks

Metastatic

Bone marrow

Genes expressed

tumor necrosis factor (TNF)

Expression markers

Tumor necrosis factor (TNF) type B, expressed; tumor necrosis factor (TNF) type A, expressed

Comments

The cells exhibit high MDR1 expression.

Recombinant tumor necrosis factor (TNF) stimulates the proliferation of SK-N-FI cells in both serum free medium and fetal bovine serum supplemented medium, but has no effect in medium without insulin.

Unpacking and storage instructions

1.      Check all containers for leakage or breakage.

2.      Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.

Complete medium

The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:

l  0.1 mM Non-Essential Amino Acids (NEAA)

l  fetal bovine serum to a final concentration of 10%

Temperature

37°C

Atmosphere

95% Air, 5% CO₂

Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

1.        Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of 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 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.        Transfer the vial contents to a 75 cm² tissue culture flask and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio). 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 complete 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).

4.        Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

Note: It is not necessary to remove the cryoprotective agent.  If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes.  Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch information.

Subculturing procedure

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1.        Remove and discard culture medium.

2.        Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.

3.        Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and ob-serve cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.

4.        Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.

5.        Add appropriate aliquots of the cell suspension to new culture vessels.

6.        Incubate cultures at 37°C.

Subculture Ratio: 1:4

Medium Renewal: 2 to 3 times a week.

Reagents for cryopreservation

Complete growth medium supplemented with 5% (v/v) DMSO.

Mycoplasma contamination

Not detected

STR profiling

Amelogenin: X,Y

CSF1PO: 10,12

D13S317: 11,14

D16S539: 11,12

D5S818: 12,13

D7S820: 8,9

TH01: 6,9.3

TPOX: 8,9

vWA: 16,17

D3S1358: 16,18

D21S11: 28,30

D18S51: 12,14

Penta_E: 11,20

Penta_D: 2.2,13

D8S1179: 11,12

FGA: 22

D19S433: 12

D2S1338: 17,18