R3 [33-10ras3]細胞

ATCC Number： CRL-2764?

細胞形態(tài)： 神經元

生長狀態(tài)： 貼壁生長

運輸方式： 凍存運輸

細胞類型： 其他細胞類型

是否是腫瘤細胞： 0

物種來源： 褐鼠

數量： 大量

Designations: R3 [33-10ras3]

Depositors: PI Patel

R3 [33-10ras3]細胞Biosafety Level: 2 [Cells contain SV40 viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Rattus norvegicus

Morphology: neuronal

Source: Cell Type: neuronal Schwann cell; immortalized with SV40 large T antigen

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Temperature Effects: Restrictive temperature: 39.5°C yes

Permissive temperature: 33°C yes

Tumorigenic: Yes

R3 [33-10ras3]細胞Comments: AntigenExp:

SV40-tsA58 large T antigen [PubMed:3049071]Oncogene:

v-Ha-ras[PubMed:3049071]Products:

Myelin-associated glycoprotein (Mag); Protein and mRNA, negative [PubMed: 12210843]Myelin basic protein (Mbp); Protein and mRNA, negative [PubMed: 12210843]Myelin protein zero (Mpz) (Charcot-Marie-Tooth neuropathy 1B); Protein and mRNA, negative [PubMed: 12210843]peripheral myelin protein 22 (Pmp22); protein, negative; mRNA, positive; [PubMed: 12210843]S100 calcium-binding protein, beta (neural) (S100b) protein, positive [PubMed: 12210843]Receptors:

Nerve growth factor receptor (Ngfr) Protein and mRNA, negative

Comments:

Primary cells were infected with the recombinant retrovirus LJ-tsSVLT containing the temperature sensitive tsA58 large SV40 T oncogene and a gene encoding G418 resistance. Clones were selected in the presence of 0.4 mg/ml G418 and super-infected with Zipras 6 containing v-Ha-ras at the permissive temperature of 33C [PubMed:3049071]. The cells proliferate at both the permissive temperature of 33C and the restrictive temperature of 39.5C. However, the growth rate is increased and growth factor requirements are reduced at the permissive temperature [PubMed: 3049071]. R3 [33-10ras3]細胞At the restrictive temperature, expression of large T antigen was not detected [PubMed: 3049071]. Expression of p21-v-Ha-ras was not detected [PubMed: 3049071].

A culture submitted to the ATCC in December of 2002 was found to be contaminated with mycoplasma. Progeny were cured by a 21-day treatment with BM Cycline.The cells were assayed for mycoplasma, by the Hoechst stain, PCR and the standard culture test, after a six-week period following treatment. All tests were negative.

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: air, 95%; carbon dioxide (CO2), 5%

Temperature: 33.0°C

Subculturing: Protocol:

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 that 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 33?C to facilitate dispersal.

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

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

Incubate cultures at 33?C.

Subcultivation Ratio: R3 [33-10ras3]細胞A subcultivation ratio of 1:4 to 1:8 is recommended

Medium Renewal: Every 2 to 3 days

Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

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

recommended serum:ATCC 30-2020

References: 64424: Hai M, et al. Comparative analysis of Schwann cell lines as model systems for myelin gene transcription studies. J. Neurosci. Res. 69: 497-508, 2002. PubMed: 12210843

71896: Ridley AJ, et al. Ras-mediated cell cycle arrest is altered by nuclear oncogenes to induce Schwann cell transformation. EMBO J. 7: 1635-1645, 1988. PubMed: 3049071