數(shù)量：大量
器官來源：眼
年限：7 days
生長狀態(tài)：貼壁生長
細(xì)胞形態(tài)：上皮樣
組織來源：retinal pigmented epithelium; retina
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ATCC Number：CRL-2240?
品系：Long-Evans
物種來源：大鼠
是否是腫瘤細(xì)胞：0
細(xì)胞類型：其他細(xì)胞類型
運(yùn)輸方式：凍存運(yùn)輸
規(guī)格：10次 Designations: RPE-J
Depositors: ?E Rodriquez-Boulan
Biosafety Level:2 [Papovavirus (cells contain SV-40 virus) ]
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Rattus norvegicus deposited as Rattus sp.
Morphology:epithelial
Source: Organ: eye
Strain: Long-Evans
Tissue: retinal pigmented epithelium; retina
Disease: normal
Cell Type: epithelialSV40 transformed
Cellular Products:SV40 T antigen
tight junction protein 1 (zona occludens 1)
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Tumorigenic:No
Age: 7 days
Comments: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 cells were transformed at 33C with the temperature-sensitive tsA SV40 virus, and cloned by limiting dilution. 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).
The cells express a transformed phenotype at the permissive temperature (33C), and a non-transformed phenotype at the non-permissive temperature (40C). They must be cultured at the permissive temperature and do not grow at 37C.
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 33C and then switched to the non-permissive temperature of 40C 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 [PubMed ID: 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 [PubMed ID: 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.
Propagation: ATCC 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%
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 33.0°C
Subculturing: Protocol:
- 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, which 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 with 5 to 10 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 37C to facilitate dispersal. - Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
- Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
- Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Place culture vessels in incubators at 33C.
Subcultivation Ratio: A subcultivation ratio of 1:3 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium with an additional 16% fetal bovine serum and 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 22464: Nabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696
