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數(shù)量：大量
運輸方式：凍存運輸
ATCC Number：CRL-10742?
器官來源：大腦
細胞類型：其他細胞類型
是否是腫瘤細胞：0
物種來源：人
年限：7 years
生長狀態(tài)：貼壁生長
細胞形態(tài)：神經(jīng)元
規(guī)格：48T Designations: HCN-2
Depositors: ?Johns Hopkins University
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Homo sapiens
Morphology:neuronal


Source: Organ: brain
Cell Type: cortical neuron;
Cellular Products:tubulin; neurofilament protein; somatostatin; cholecystokinin-8
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.
Applications:The cells are negative for glial fibrillary acidic protein (GFAP) and myelin basis protein (MBP).
The cells stain positively for a number of neuronal markers including neurofilament protein, neuron specific enolase (NSE).
The HCN-2 cell line was derived from cortical tissue removed from a patient undergoing hemispherectomy for intractable seizures associated with Rasmussen's encephalitis.
They are also positive for tubulin, vimentin, somatostatin (SST), glutamate, gamma aminobutyric acid (GABA), cholecystokinin - 8 (CCK-8) and vasoactive intestinal peptide (VIP).
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HCN-2 cells can be induced to differentiate when cultured with a mixture of nerve growth factor (NGF), dibutyryl cyclic adenosine monophosphate (cAMP) and 1-isobutyl-3-methylxanthine (IBMX).
DNA Profile (STR):Amelogenin: X
CSF1PO: 10,15
D13S317: 11,12
D16S539: 8,11
D5S818: 13
D7S820: 9,11
THO1: 6,9.3
TPOX: 8
vWA: 14,20
Age: 7 years
Gender: female
Comments:HCN-2 cells can be induced to differentiate when cultured with a mixture of nerve growth factor (NGF), dibutyryl cyclic adenosine monophosphate (cAMP) and 1-isobutyl-3-methylxanthine (IBMX). Differentiation is accompanied by mature morphology and slowing of growth (doubling time greater than 120 hours). The growth rate of HCN-2 cells is stimulated by treatment with phorbol esters. A similar line (HCN-1A) is available as ATCC CRL-10442.
The HCN-2 cell line was derived from cortical tissue removed from a patient undergoing hemispherectomy for intractable seizures associated with Rasmussen's encephalitis.
The cells stain positively for a number of neuronal markers including neurofilament protein, neuron specific enolase (NSE).
They are also positive for tubulin, vimentin, somatostatin (SST), glutamate, gamma aminobutyric acid (GABA), cholecystokinin - 8 (CCK-8) and vasoactive intestinal peptide (VIP).
The cells are negative for glial fibrillary acidic protein (GFAP) and myelin basis protein (MBP).
CRL-10742 has been shown to senesce at approximately passage 21. Current distribution stocks are prepared with only 4-5 Population Doublings (PDLs) remaining under recommended culture conditions after cryopreservation.
Propagation: ATCC complete growth medium: Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 4.5 g/L glucose, 90%; fetal bovine serum, 10%
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0℃
Subculturing: Protocol: NOTE: This cell line is extremely slow growing. May not be able to subculture for 12 to 14 days after recovery. Cultures do not reach 100% confluence (about 80-90%). A change in the fetal bovine serum may help increase the growth rate. Subculture at no higher than a 1:3 ratio every 10 to 12 days when cultures are about 90% confluent.

1. Remove and discard culture medium.
2. 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.
3. 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.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
6. Incubate cultures at 37?C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 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: 22022: Ronnett GV, et al. Human neuronal cell line. US Patent 5,196,315 dated Mar 23 1993