| 產(chǎn)品名稱 |
MLE 12 |
| 商品貨號 |
B165163 |
| Organism |
Mus musculus, transgenic, mouse, transgenic |
| Tissue |
lung |
| Cell Type |
epithelial; SV40 transformed |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Papovavirus]
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. |
| Age |
5 month old |
| Gender |
female |
| Strain |
FVB/N |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
This line was established in 1992 by Kathryn A. Wikenheiser from pulmonary tumors in a mouse transgenic for the SV40 large T antigen under the control of the promoter region of the human surfactant protein C gene. |
| Clinical Data |
female
5 months |
| Genes Expressed |
lung surfactant proteins B and C (SP-B, SP-C) |
| Tumorigenic |
Yes |
| Effects |
Yes, tumors were generated within 6 to 9 months in nude mice |
| Comments |
The cells express the mRNA for large T antigen.
Lung surfactant proteins B and C were detected.
The cells secrete phospholipids in response to phorbol esters and ATP but not in response to forskolin. |
| Complete Growth Medium |
HITES medium supplemented with 2% fetal bovine serum
HITES medium with 2% fetal bovine serum is formulated at the ATCC as follows:
- Dulbecco's medium : Ham's F12, 50:50 mix (ATCC® 30-2006)
- Insulin 0.005 mg/ml
- Transferrin 0.01 mg/ml
- Sodium selenite 30 nM
- Hydrocortisone 10 nM
- β-estradiol 10 nM
- HEPES 10 mM
- L-glutamine (ATCC® 30-2214) 2 mM (in addition to that in the base medium)
- Fetal bovine serum (ATCC® 30-2020) 2%
|
| Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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.
Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:10 is recommended
Medium Renewal: Twice per week |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
| Population Doubling Time |
7 to 9 hrs |
| Name of Depositor |
JA Whitsett |
| Deposited As |
mouse, transgenic |
| Year of Origin |
1992 |
| References |
Wikenheiser KA, et al. Simian virus 40 large T antigen directed by transcriptional elements of the human surfactant protein C gene produces pulmonary adenocarcinomas in transgenic mice. Cancer Res. 52: 5342-5352, 1992. PubMed: 1394139
Wikenheiser KA, et al. Production of immortalized distal respiratory epithelial cell lines from surfactant protein C/simian virus 40 large tumor antigen transgenic mice. Proc. Natl. Acad. Sci. USA 90: 11029-11033, 1993. PubMed: 8248207
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