RN33B細(xì)胞

年限： 12.5 days gestation embryo

ATCC Number： CRL-2825?

細(xì)胞形態(tài)： 其他

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 褐鼠

生長狀態(tài)： 貼壁生長

器官來源： 大腦

數(shù)量： 大量

品系： Sprague-Dawley

組織來源： medullary raphe nucleus

運(yùn)輸方式： 凍存運(yùn)輸

Designations: RN33B

Depositors: SR Whittemore

RN33B細(xì)胞Biosafety Level: 2 [cells containing SV40 viral DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Rattus norvegicus

Morphology: spindle-shaped at 33C; neurite-like at 37C

Source: Strain: Sprague-Dawley

Organ: brain

Tissue: medullary raphe nucleus

Cell Type: neuronalSV40 large T antigen transfected

Cellular Products: enolase

nestin

neurofilament

vimentin

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. RN33B細(xì)胞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: February, 1988

Receptors: nerve growth factor (NGF), expressed

Temperature Effects: Restrictive temperature: 37 to 39°C cells differentiate into neurons

Permissive temperature: 33°C cells proliferate

Y

Age: 12.5 days gestation embryo

Comments: RN33B is an neuronal cell line derived from medullary raphe cells by infection with a retrovirus encoding the temperature-sensitive mutant of SV40 large T antigen. The cells were cloned by serial dilution.At the permissive temperature of 33C, RN33B cells divide and express SV40 T antigen, vimentin, nestin, diffuse neuron-specific enolase, and low and medium molecular weight neurofilament immunoreactivities.At the non-permissive temperature of 37C to 39C SV40 T antigen expression is markedly decreased and RN33B cells cease mitotic activity and differentiate with phase bright cell bodies and 'neuritic-like' processes.RN33B細(xì)胞Differentiated RN33B cells express enhanced neuronal-specific protein expression but do not synthesize astrocytic or oligodendrocytic-specific proteins. Moreover, differentiated RN33B cells returned to 33C re-express T antigen, but do not de-differentiate or begin dividing. Immunohistochemical and Northern blot analysis revealed high levels of low affinity NGF receptor protein and mRNA in differentiated RN33B cells. PCR analysis demonstrated the presence of trkB, but not trkA or trkC, mRNA in both undifferentiated and differentiated RN33B cells. The cells can be used as a model of neuronal differentiation in vitro.

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No. 30-2006. 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 33C 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.

An inoculum of 5 X 10(3) to 8 X 10(3) viable cells/sq. cm is recommended.

RN33B細(xì)胞Incubate cultures at 33C.

Interval: Maintain cultures at a cell concentration between 3 X 10(4) and 1 X 10(5) cells/sq. cm

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended

Medium Renewal: Two to three times weekly

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 20 hours

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006

recommended serum:ATCC 30-2020

References: 89616: Whittemore SR, White LA. Target regulation of neuronal differentiation in a temperature-sensitive cell line derived from medullary raphe. Brain Res. 615: 27-40, 1993. PubMed: 8364724

89617: Shihabuddin LS, et al. The ***** CNS retains the potential to direct region-specific differentiation of a transplanted neuronal precursor cell line. J. Neurosci. 15: 6666-6678, 1995. PubMed: 7472427

89618: Wojciechowski AB, et al. Long-term survival and glial differentiation of the brain-derived precursor cell line RN33B after subretinal transplantation to ***** normal rats. Stem Cells 20: 163-173, 2002. PubMed: 11897873

89619: Warfvinge K, et al. Retinal integration of grafts of brain-derived precursor cell lines implanted subretinally into *****, normal rats. Exp. Neurol. 169: 1-12, 2001. PubMed: 11312552