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M059K  

神经胶质瘤细胞M059K

ATCC® Number: CRL-2365™    
Designations: M059K
Depositors:  J Allalunis-Turner, RS Day
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens (human)
Morphology: fibroblast

Source: Organ: brain
Disease: malignant glioblastoma; glioma
Cell Type: glial cell;
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.
Tumorigenic: Yes
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 10,12
D13S317: 14
D16S539: 10,12
D5S818: 11,12
D7S820: 10
THO1: 9.3
TPOX: 8
vWA: 17
Cytogenetic Analysis: Number of cells examined = 59; Modal Chromosome Number = 75 with a range of 65 to 79; Polyploidy Rate = 22%
Age: 33 years
Gender: male
Comments: M059K cells were isolated from a tumor specimen from a 33 year-old male patient with untreated glioblastoma. These cells were isolated concurrently from the same tumor specimen as M059J (see ATCC CRL-2366). M059K cells express normal levels of DNA-dependent protein kinase while M059J cells lack DNA-dependent protein kinase activity.M059K cells are approximately 30-fold less sensitive to ionizing radiation than M059J cells.M059K cells are less sensitive than M059J cells to the cytotoxic effects of bleomycin, N, N-bis(2-chloroethyl)-N-nitrosourea and nitrogen mustard.M059K cells are DNA double strand break repair proficient.Both cell lines are negative for glial fibrillary acidic acid (GFAP).Together these cell lines provide useful model systems in which to study the role of DNA protein kinase in cellular and molecular processes involving DNA damage recognition and repair.
Propagation: ATCC 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.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
  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 which 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 37C 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 37C.

Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor temperature
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006
recommended serum:ATCC 30-2020
derived from same individual:ATCC CRL-2366
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 33940: 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
33942: 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
38596: 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
38598: 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
38599: 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