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微信小章| 产品名称： | RPE-J |
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| 商品货号： | TS210996 |
| 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)
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. |
| 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.xa0Ref 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.Ref 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.
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| Complete Growth Medium： | Dulbeccos modified Eagles 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%
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| 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.
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,xa05th edition, published byxa0Wiley-Liss, N.Y., 2005.
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| 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 |
| Deposited As： | Rattus sp. |
| References： | Nabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696 |
Nabi IR, et al. Immortalization of polarized rat retinal pigment epithelium. J. Cell Sci. 104: 37-49, 1993. PubMed: 8383696