sNF96.2細(xì)胞

運(yùn)輸方式： 凍存運(yùn)輸

ATCC Number： CRL-2884?

相關(guān)**： 其他**

年限： 28 year old

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 人

數(shù)量： 大量

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

細(xì)胞形態(tài)： 其他

Designations: sNF96.2

Depositors: DF Muir, MR Wallace

sNF96.2細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: Schwann cell-like

Source: Disease: neurofibrmatosis type 1 (NF1)

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Isolation: Isolation date: 1996

DNA Profile (STR): Amelogenin: X

CSF1PO: 12

D13S317: 10

D16S539: 11

sNF96.2細(xì)胞D5S818: 11

D7S820: 10,11

THO1: 6

TPOX: 11

vWA: 17,19

Age: 28 year old

Gender: male

Comments: The sNF96.2 cell line was derived from a recurrent mass associated with nerve and diagnosed as MPNST (Malignant Peripheral Nerve Sheath Tumor) in a patient meeting NF1 diagnostic criteria. The cells were derived from numerous passages of primary tumor material in culture until they were a homogenous Schwann cell-like population which displayed a clonal morphology immunopositive for both cytoplasmic Schwann cell markers S100 and p75. The line has an abnormal karyotype and complete LOH (Loss of Heterozygocity) with no detection of the remaining NF1 allele. Western immunoblotting showed no full-length neurofibromin protein.

Propagation: sNF96.2細(xì)胞ATCC complete growth 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: fetal bovine serum to a final concentration of 10%.

Atmosphere: 5% CO2 in air recommended

Temperature: 37.0°C

Subculturing: Protocol:

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

Remove and discard culture medium.

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

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe 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.

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

Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 5 X 10(3) to 7 X 10(3) viable cells/sq. cm is recommended.

Incubate cultures at 37C. Subculture when cell concentration is between 1 X 10(4) and 2 X 10(4) cells/sq. cm.

Subcultivation Ratio: sNF96.2細(xì)胞A subcultivation ratio of 1:3 to 1:4 twice weekly is recommended

Medium Renewal: Every 2 to 3 days

Preservation: Freeze medium: culture medium,90%; DMSO,10%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: about 33 hours

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

recommended serum:ATCC 30-2020

0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101

Cell culture tested DMSO:ATCC 4-X

Erythrosin B vital stain solution:ATCC 30-2404

Trypan Blue vital stain solution:ATCC 30-2402

sNF96.2細(xì)胞References: 90346: Fieber LA, et al. Delayed rectifier K currents in NF1 Schwann cells. Pharmacological block inhibits proliferation. Neurobiol. Dis. 13: 136-146, 2003. PubMed: 12828937

90347: Li Y, et al. Notch and Schwann cell transformation. Oncogene 23: 1146-1152, 2004. PubMed: 14762442