RLE-6TN (ATCC® CRL-2300)

Organism: Rattus norvegicus, rat  /  Cell Type: alveolar type II; SV40 transfecte  /  Tissue: lung  / 

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Organism Rattus norvegicus, rat
Tissue
lung
Cell Type alveolar type II; SV40 transfecte
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 Cells contain Papovavirus
Age 56 day old
Gender male
Strain Fischer 344 (F344)
Applications
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.
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.
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
Expression of the SV40-T antigen was negative by nuclear immunostaining and by PCR, indicating these cells were derived by a spontaneous immortalization.
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.
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,However, expression of the SV40-T antigen was negative by nuclear immunostaining and by PCR, indicating these cells were derived by a spontaneous immortalization.,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
The endothelial nature of these cells was confirmed by the observed expression of von Willebrand factor and uptake of fluorescently labeled low density lipoprotein (LDL).
Tumorigenic No
Effects
No, not tumorigenic in nude mice
Comments
At passage 5, the alveolar Type II cells were transfected with SV40 (pRSV-T DNA) by lipofection.
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.
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
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 that 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 37°C 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 37°C.
.
Subcultivation Ratio: A subcultivation ratio of 1:5 is recommended
Medium Renewal: Every 2 to 3 days
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
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