| 產品名稱 |
LN-229 |
| 商品貨號 |
B164994 |
| Organism |
Homo sapiens, human |
| Tissue |
brain/right frontal parieto-occipital cortex |
| Cell Type |
Glioblastoma |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
glioblastoma |
| Age |
60 years |
| Gender |
female |
| Ethnicity |
White |
| Applications |
This cell line is used in studies on apoptosis.
|
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The LN-229 cell line was established in 1979 from cells taken from a patient with right frontal parieto-occipital glioblastoma [PubMed. 10416987]. |
| Clinical Data |
60 years
female
White |
| Oncogene |
p53 + (mutated, CCT (Pro) --> CTT (Leu) mutation at codon 98), PTEN + (wild type), p16 - (deleted), p14ARF - (deleted) |
| Tumorigenic |
Yes |
| Effects |
Yes, forms tumors in nude mice |
| Comments |
The cells exhibit mutated p53 (TP53) and possible homozygous deletions in the p16 and p14ARF tumor suppressor genes. They have a wild-type PTEN gene.
Stimulation of the cells with Fas ligand lead to apoptotic cell death within 16 hours. The cells were also killed by puromycin in a dose dependent manner.
Bcl-2 protects these cells from Fas ligand-induced cell death but was shown to have only a small protective effect on puromycin-induced apoptosis.
|
| 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 5%.
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: Subculture at 80% confluence. Cells pile up and do not become completely confluent.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53% (w/v) 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 37°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 37°C
Subculture Ratio: 1:4 to 1:6
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994
|
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C |
| STR Profile |
Amelogenin: X CSF1PO: 12 D13S317: 10,11 D16S539: 12 D5S818: 11,12 D7S820: 8,11 THO1: 9.3 TPOX: 8 vWA: 16,19 |
| Name of Depositor |
N de Tribolet |
| Deposited As |
human |
| Year of Origin |
1979 |
| References |
Diserens AC, et al. Characterization of an established human malignant glioma cell line: LN-18. Acta Neuropathol. 53: 21-28, 1981. PubMed: 7211194
Ishii N, et al. Frequent co-alterations of TP53, p16/CDKN2A, p14ARF, PTEN tumor suppressor genes in human glioma cell lines. Brain Pathol. 9: 469-479, 1999. PubMed: 10416987
Schlapbach R, Fontana A. Differential activity of bcl-2 and ICE enzyme family protease inhibitors on Fas and puromycin-induced apoptosis of glioma cells.. Biochim. Biophys. Acta 1359: 174-180, 1997. PubMed: 9409814
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