生长状态:贴壁生长
细胞形态:神经元
细胞类型:其他细胞类型
品系:BD1X
年限:4 to 10 months
是否是肿瘤细胞:0
物种来源:褐鼠
ATCC Number:CRL-2754?
运输方式:冻存运输
相关疾病:神经母细胞瘤
器官来源:其他
数量:大量
规格:0.1ml Designations: B35
Depositors: ?P Maness
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism:
Rattus norvegicus
Morphology:neuronal
Source:
Organ: central nervous system (CNS)
Strain: BD1X
Disease: neuroblastoma
Cell Type: neuronal neuroblast; nitrosoethylurea (NEU) induced
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.
Age: 4 to 10 months
Comments:Rats were inoculated with N-nitrosoethylurea (NEU) 15 days after conception. Tumors found in the central nervous system (CNS) 4 to 10 months after birth were excised, minced, adapted to culture and cloned [PubMed: 4151463]. B35 cells can be stimulated to differentiate in the presence of dibutyryl cyclic AMP (cAMP) or by serum deprivation. They are easily transfected with plasmid DNA. The cells retain glutamic acid decarboxylase (GAD) and choline acetyltransferase activities; express gamma aminobutyric acid (GABA). The cells are negative for S100 (S-100) protein [PubMed: 4151463]. The cells are positive for neuron specific enolase [PubMed: 6722796].The cells also may be used to study the metabolism and physiology of nervous tissue and the pathology of nervous disorders. A culture submitted to the ATCC in October 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%.
Temperature: 37.0°C
Subculturing:
Protocol: Subculture before confluency
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Remove and discard culture medium.
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Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
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Incubate cultures at 37?C.
Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Preservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
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: 61205: Schubert D, et al. Clonal cell lines from the rat central nervous system. Nature 249: 224-227, 1974. PubMed: 4151463
61314: Vinores SA, et al. Immunoradiometric and immunohistochemical demonstration of neuron-specific enolase in experimental rat gliomas. Cancer Res. 44: 2595-2599, 1984. PubMed: 6722796
88865: Otey CA, et al. B35 neuroblastoma cells: an easily transfected, cultured cell model of central nervous system neurons. Methods Cell Biol. : 287-304, 2003. PubMed: 12884695