LN-18 (ATCC® CRL-2610)

Organism: Homo sapiens, human  /  Tissue: brain/cerebrum; right temporal lobe  /  Disease: grade IV,glioblastoma; glioma

Organism Homo sapiens, human
Tissue brain/cerebrum; right temporal lobe
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease grade IV,glioblastoma; glioma
Age 65 years
Gender male
Ethnicity White
Applications
This cell line is used in studies on apoptosis.
Storage Conditions liquid nitrogen vapor phase
Derivation
The LN-18 cell line was established in 1976 from cells taken from a patient with a right temporal lobe glioma. ­
Clinical Data
65 years
male
White
Antigen Expression
HLA A2, A9, B5, BW35, DRW3
Oncogene p53 + (mutated, TGT (Cys) --> TCT (Ser) mutation at codon 238), PTEN + (wild type), p16 - (deleted), p14ARF - (deleted) RefFlaman JM, et al. A simple p53 functional assay for screening cell lines, blood, and tumors. Proc. Natl. Acad. Sci. USA 92: 3963-3967, 1995. PubMed: 7732013
Genes Expressed
fibronectin
Cellular Products
fibronectin
Tumorigenic Yes
Effects
Yes, forms tumors in nude mice
Comments

The cells are negative for glial fibrillary acidic proteins and S100 (S-100) protein.

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.

  1. Remove and discard culture medium.
  2. 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.
  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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. 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, 90%; additional fetal bovine serum, 5%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 12
D13S317: 12,13
D16S539: 11,13
D5S818: 11,13
D7S820: 8,10
THO1: 9
TPOX: 8
vWA: 17,18
Name of Depositor N de Tribolet
Year of Origin 1976
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

Flaman JM, et al. A simple p53 functional assay for screening cell lines, blood, and tumors. Proc. Natl. Acad. Sci. USA 92: 3963-3967, 1995. PubMed: 7732013

Flaman JM, et al. A simple p53 functional assay for screening cell lines, blood, and tumors. Proc. Natl. Acad. Sci. USA 92: 3963-3967, 1995. PubMed: 7732013

Basic Documentation
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

Flaman JM, et al. A simple p53 functional assay for screening cell lines, blood, and tumors. Proc. Natl. Acad. Sci. USA 92: 3963-3967, 1995. PubMed: 7732013

Flaman JM, et al. A simple p53 functional assay for screening cell lines, blood, and tumors. Proc. Natl. Acad. Sci. USA 92: 3963-3967, 1995. PubMed: 7732013