RPE-J (ATCC® CRL-2240)

Organism: Rattus norvegicus, rat  /  Cell Type: epithelial SV40 transformed  /  Tissue: eye/retinal pigmented epithelium; retina  /  Disease: normal

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Organism Rattus norvegicus, rat
Tissue eye/retinal pigmented epithelium; retina
Cell Type epithelial SV40 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Papovavirus (cells contain SV-40 virus)]
Disease normal
Age 7 days
Strain Long-Evans
Storage Conditions liquid nitrogen vapor phase
Derivation

RPE-J is a retinal pigment epithelial (RPE) cell line derived from primary cultures of RPE cells taken from 7-day-old Long-Evans rats.

The RPE-J clone was selected for an epithelioid morphology and for expression of circumferential staining with antibody against the tight junction protein 1, (Tjp1, ZO-1).

Genes Expressed
SV40 T antigen,tight junction protein 1 (zona occludens 1)
Cellular Products
SV40 T antigen
tight junction protein 1 (zona occludens 1)
Tumorigenic No
Effects
No, in immunosuppressed mice
Yes, in semisolid medium (colony formation)
Comments

The cells express a transformed phenotype at the permissive temperature (33°C), and a non-transformed phenotype at the non-permissive temperature (40°C). They must be cultured at the permissive temperature and do not grow at 37°C.

When RPE-J cells are grown on nitrocellulose filters coated with a thin layer of Matrigel in the presence of 10(-8) M retinoic acid for 6 days at 33°C and then switched to the non-permissive temperature of 40°C for 33 to 36 hours, they acquire a differentiated polarized RPE phenotype. Under these conditions, RPE-J cells exhibit circumferential staining for the tight-junction protein ZO-1 and acquire a transepithelial resistance of 350 ohms/cm2. RefNabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696

RPE-J is the only established RPE cell line that maintains epithelial cell surface polarity. The cells retain many properties of RPE including expression of the rat RPE marker RET-PE2 and the ability to phagocytose latex beads.RefNabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696

A culture submitted to the ATCC in July 1995 was found to be contaminated with Mycoplasma hyorhinis and was cured by a 21-day treatment with BM Cycline. The cells were assayed for mycoplasma by the Hoechst stain and the standard culture test over a six-week period following treatment and all tests were negative.

RPE-J is a retinal pigment epithelial (RPE) cell line derived from primary cultures of RPE cells taken from 7-day-old Long-Evans rats.

Complete Growth Medium Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5g/L sodium bicarbonate and 4.5 g/L glucose and supplemented with 0.1 mM non-essential amino acids, 96%; fetal bovine serum, 4%
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.

  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. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium.  Add appropriate aliquots of cell suspension to new culture vessels.
  7. Place culture vessels in incubators at 33°C.

Subculture Ratio: 1:3
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.

Cryopreservation
Freeze medium: Complete growth medium with an additional 16% fetal bovine serum and 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 33°C
Name of Depositor E Rodriquez-Boulan
References

Nabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696

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

Nabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696