產(chǎn)品名稱 |
M059K |
商品貨號 |
B165031 |
Organism |
Homo sapiens, human |
Tissue |
brain |
Cell Type |
glial cell |
Product Format |
frozen |
Morphology |
fibroblast |
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 |
malignant glioblastoma; glioma |
Age |
33 years |
Gender |
male |
Applications |
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. |
Storage Conditions |
liquid nitrogen vapor phase |
Karyotype |
Number of cells examined = 59; Modal Chromosome Number = 75 with a range of 65 to 79; Polyploidy Rate = 22% |
Images |
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Derivation |
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). - |
Clinical Data |
male
33 years |
Tumorigenic |
Yes |
Effects |
Yes, forms tumors in SCID mice |
Comments |
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. |
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.
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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.
- Remove and discard culture medium.
- 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.
- 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 10 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: 1:6 to 1:8
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
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Cryopreservation |
Freeze Medium: Complete growth medium 95%; DMSO, 5%
Storage Temperature: Liquid nitrogen vapor temperature |
Culture Conditions |
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
STR Profile |
Amelogenin: X,Y CSF1PO: 10,12 D13S317: 14 D16S539: 10,12 D5S818: 11,12 D7S820: 10 THO1: 9.3 TPOX: 8 vWA: 17 |
Name of Depositor |
J Allalunis-Turner, RS Day |
Deposited As |
human |
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
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