M059J (ATCC® CRL-2366)

Organism: Homo sapiens, human  /  Cell Type: glial cell  /  Tissue: brain  /  Disease: malignant glioblastoma; glioma

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Organism Homo sapiens, human
Tissue brain
Cell Type glial cell
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease malignant glioblastoma; glioma
Age 33 years
Gender male
Applications
Together, M059K and M059J provide a useful model system in which to study the role of DNA protein kinase in cellular and molecular processes involving DNA damage recognition and repair
Storage Conditions liquid nitrogen vapor temperature
Karyotype aneuploid; Y chromosome is present
Derivation
M059J cells were isolated from a tumor specimen taken from a 33 year old male with untreated malignant glioblastoma
The cells were isolated concurrently from the same tumor specimen as M059K (see CRL-2365).
Clinical Data
male
33 years
Comments
M059J cells lack DNA-dependent protein kinase activity, while M059K cells express normal levels of DNA-dependent protein kinase.
M059J cells are approximately 30-fold more sensitive to ionizing radiation than M059K cells.
M059J cells are more sensitive than M059K cells to the cytotoxic effects of bleomycin, N,N-bis(2-choroethyl)-N-nitrosourea and nitrogen mustard.
M059J cells are deficient in repair of DNA double strand breaks.
The cells are negative for glial fibrillary acidic protein (GFAP).
Complete Growth Medium These cells are grown in a medium containing a 1:1 mixture of Dulbecco's Modified Eagle's Medium and Ham's F12 medium with 2.5 mM L-glutamine adjusted to contain 15 mM HEPES, 0.5 mM sodium pyruvate, and 1.2 g/L sodium bicarbonate supplemented with 0.05 mM non-essential amino acids and 10% fetal bovine serum.
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 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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C

Subculture Ratio: 1:6 to 1:8
Medium Renewal: Every 2 to 3 days.
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 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor temperature
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 10,12
D13S317: 14
D16S539: 10,12
D5S818: 11,12
D7S820: 10,12
THO1: 9.3
TPOX: 8
vWA: 17
Name of Depositor J Allalunis-Turner, RS Day
References

Allalunis-Turner MJ, et al. Isolation of two lines from a human malignant glioma specimen differing in sensitivity to radiation and chemotherapeutic drugs. Radiat. Res. 134: 349-354, 1993. PubMed: 8316628

Lees-Miller SP, et al. Absence of p350 subunit of DNA activated protein kinase from a radiosensitive human cell line. Science 267: 1183-1185, 1995. PubMed: 7855602

Allalunis-Turner J, et al. Intact G2-phase checkpoint in cells of a human cell line lacking DNA-dependent protein kinase activity. Radiat. Res. 147: 284-287, 1997. PubMed: 9052673

Allalunis-Turner MJ, et al. Radiation-induced DNA damage and repair in cells of a radiosensitive human malignant glioma cell line. Radiat. Res. 144: 288-293, 1995. PubMed: 7494872

Wang J, et al. Radiation-induced damage in two human glioma cell lines as measured by the nucleoid assay. Anticancer Res. 17: 4615-4618, 1997. PubMed: 9494578

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Allalunis-Turner MJ, et al. Isolation of two lines from a human malignant glioma specimen differing in sensitivity to radiation and chemotherapeutic drugs. Radiat. Res. 134: 349-354, 1993. PubMed: 8316628

Lees-Miller SP, et al. Absence of p350 subunit of DNA activated protein kinase from a radiosensitive human cell line. Science 267: 1183-1185, 1995. PubMed: 7855602

Allalunis-Turner J, et al. Intact G2-phase checkpoint in cells of a human cell line lacking DNA-dependent protein kinase activity. Radiat. Res. 147: 284-287, 1997. PubMed: 9052673

Allalunis-Turner MJ, et al. Radiation-induced DNA damage and repair in cells of a radiosensitive human malignant glioma cell line. Radiat. Res. 144: 288-293, 1995. PubMed: 7494872

Wang J, et al. Radiation-induced damage in two human glioma cell lines as measured by the nucleoid assay. Anticancer Res. 17: 4615-4618, 1997. PubMed: 9494578