| 產(chǎn)品名稱 |
RLE-6TN |
| 商品貨號(hào) |
B165609 |
| Organism |
Rattus norvegicus, rat |
| Tissue |
lung |
| Cell Type |
alveolar type II; SV40 transfected |
| 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 |
56 day old |
| Gender |
male |
| Strain |
Fischer 344 (F344) |
| Storage Conditions |
liquid nitrogen vapor phase |
| Karyotype |
Cells are reported to remain near diploid and karyotypically stable from passage 19-70 with 50% or more of the cells containing 42 chromosomes. At passage 37, there was a translocation between chromosome 1 and 15 which results in trisomy of the q arm of chromosome 1. |
| Derivation |
The RLE-6TN (rat lung epithelial-T-antigen negative) cell line was derived from alveolar Type II cells isolated from a 56 day old male F344 rat using airway perfusion with a pronase solution. Expression of the SV40-T antigen was negative by nuclear immunostaining and by PCR, indicating these cells were derived by a spontaneous immortalization. |
| Clinical Data |
male The RLE-6TN (rat lung epithelial-T-antigen negative) cell line was derived from alveolar Type II cells isolated from a 56 day old male F344 rat using airway perfusion with a pronase solution. |
| Genes Expressed |
cytokeratin 8 and 19 |
| Cellular Products |
cytokeratin 8 and 19 |
| Tumorigenic |
No |
| Effects |
No, not tumorigenic in nude mice |
| Comments |
The cell line exhibits characteristics of alveolar Type II cells such as lipid-containing inclusion bodies (phosphine 3R staining and electron microscopy) and expression of cytokeratin 8 and 19; the cells do not express alkaline phosphatase activity.
Expression of several chemotactic cytokines by RLE-6TN cells was reported to be similar to that of primary cultures of alveolar Type II cells.
Although these cells appear negative for the SV40 antigen, they should be handled as potentially biohazardous material under at least Biosafety Level 2 containment.
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| Complete Growth Medium |
Ham's F12 medium with 2 mM L-glutamine supplemented with 0.01 mg/ml bovine pituitary extract, 0.005 mg/ml insulin, 2.5 ng/ml insulin-like growth factor, 0.00125 mg/ml transferrin, and 2.5 ng/ml EGF, 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: 1:5
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 supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
KE Driscoll |
| Passage History |
At passage 5, the alveolar Type II cells were transfected with SV40 (pRSV-T DNA) by lipofection. |
| References |
Driscoll KE, et al. Establishment of immortalized alveolar type II epithelial cell lines from adult rats. In Vitro Cell. Dev. Biol. Anim. 31: 516-527, 1995. PubMed: 8528500
Driscoll KE, et al. Alpha-quartz-induced chemokine expression by rat lung epithelial cells: effects of in vivo and in vitro particle exposure. Am. J. Pathol. 149: 1627-1637, 1996. PubMed: 8909252
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