| 產品名稱 |
9L/lacZ |
| 商品貨號 |
B163858 |
| Organism |
Rattus norvegicus, rat |
| 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 |
gliosarcoma |
| Applications |
This cell line is one of few models that permit quantitative analysis of microscopic tumor in the brain.
The tumor mimics important features of human brain tumor growth and spread. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The 9L/lacZ cell line was developed in 1989 from the 9L cell line (rat nitrosourea induced gliosarcoma cell line).
9L cells were infected with the BAG replication deficient retroviral vector carrying the E. coli lacZ gene encoding beta-gal and the Tn5 neomycin gene, which confers resistance to G418.
The cells were cultured in G418 for 14 days, cloned, and evaluated for beta-gal production.
9L/lacZ produced high levels of the enzyme, and was selected for study. |
| Genes Expressed |
beta galactosidase (beta-gal) |
| Tumorigenic |
Yes |
| Effects |
Yes, forms tumors in the brains of CD Fischer 344 rats |
| Comments |
The cells constitutively express the lacZ reporter gene product, E. coli derived beta-gal, as revealed on tissue sections by histochemical stain, and single tumor cells can be identified.
Lymphocytes and other responding cells can be identified by double labeling with antibodies on the same slide.
The contrast between stained cells and background facilitates image analysis.
The beta-gal expression is very stable, but cells may need to be re-cloned after months of growth in culture. |
| Complete Growth Medium |
Dulbecco's modified Eagle's medium with 4.5 g/L glucose, and 1 mM sodium pyruvate 90%; fetal bovine serum, 10%
|
| 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 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 1:8
Medium Renewal: 2 to 3 times a week.
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 |
Culture medium, 95%; DMSO, 5% |
| Name of Depositor |
LA Lampson |
| Deposited As |
Rattus sp. |
| References |
Lampson LA, et al. Interactions between leucocytes and individual brain tumor cells in the rat brain. J. Neuro-Oncol. 7: S17, 1990.
Lampson LA, et al. Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ. Cancer Res. 53: 176-182, 1993. PubMed: 8416743
Lampson LA, et al. Disseminating tumor cells and their interactions with leukocytes visualized in the brain. Cancer Res. 52: 1018-1025, 1992. PubMed: 1737331
Dutta T, et al. Robust ability of IFN-gamma to upregulate class II MHC antigen expression in tumor bearing rat brains. J. Neuro-Oncol. 64: 31-44, 2003. PubMed: 12952284
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