運輸方式：凍存運輸
ATCC Number：CRL-2884?
相關(guān)疾?。浩渌膊?br />
年限：28 year old
是否是腫瘤細胞：0
物種來源：人
數(shù)量：大量
生長狀態(tài)：貼壁生長
細胞形態(tài)：其他
規(guī)格：1ml Designations: sNF96.2
Depositors: ?DF Muir, MR Wallace
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)
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: 1996
DNA Profile (STR):Amelogenin: X
CSF1PO: 12
D13S317: 10
D16S539: 11
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: 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: 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
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
